Optical measurement device for reaction vessel and method therefor

ABSTRACT

An optical measurement device is provided. The device includes first and second optical fibers; first and second reaction vessels, and a light guide stage coupled to the first and second optical fibers. The light guide stage is driven to simultaneously optically connect the first and second optical fibers with the first and second reaction vessels. The device includes a measurement device for receiving emissions from the first and second reaction vessels, and a connecting end arranging body that supports the first and second optical fibers along a path. The arranging body is driven along the path between a first position, in which the first optical fiber is optically connected with the measurement device so that light is transmittable from the first reaction vessel, and a second position, in which the second optical fiber is optically connected with the measurement device so that light is transmittable from the second reaction vessel.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application is a continuation of U.S. patent applicationSer. No. 16/189,663, filed Nov. 13, 2018, which is a continuation ofU.S. patent application Ser. No. 14/117,785, filed Jan. 31, 2014, whichis a 371 national phase of international patent application numberPCT/JP2012/062550, filed May 16, 2012, which claims priority to Japanesepatent application number 2011-109918, filed May 16, 2011. All of whichapplications are incorporated herein by reference, in entirety.

FIELD

The present invention relates to an optical measurement device for areaction vessel, and a method therefor.

BACKGROUND

At the time reactions such as amplification of nucleic acids (DNA, RNA,and the like) and the fragments thereof (oligonucleotides, nucleotides,and the like) are performed, in tests that require quantitativeness,such as the analysis of gene expression levels, it becomes necessary toperform the amplification such that the ratio of the relative amounts ofthe respective nucleic acids can be known. Consequently, by using thereal-time PCR method, and by using a device provided with a thermalcycler and a fluorescence spectrophotometer, analysis by electrophoresisis made unnecessary as a result of the generation process of the DNAamplification products in PCR being detected and analyzed in real time.Furthermore, as a DNA amplification method that performs amplificationwhile maintaining the quantitativeness with respect to the ratio of therelative amounts of the DNA or RNA contained in the sample beforeamplification, the SPIA (Single Primer Isothermal Amplification) methodis used. In the SPIA method, the linear DNA amplification methodresulting from an isothermal reaction utilizing DNA/RNA chimera primer,DNA polymerase, and RNaseH has become used.

In a case where processing such as nucleic acid amplification, andmeasurements thereof are performed, conventionally, the target nucleicacid is separated and extracted from the sample by using a filter bymeans of a manual method, by using magnetic particles and adsorption onan inner wall of a vessel or a pipette tip by means of a magnetic field,or by using a centrifuge. The separated and extracted target compound istransferred and introduced into a reaction vessel together with areaction solution by a manual method and the like, and upon sealing ofthe reaction vessel using a manual method and the like, at the timereactions are performed using a temperature control device forreactions, optical measurements are performed with respect to thereaction vessel using a light measuring device (Patent Document 1).

In a case where the processing is executed by a manual method, a largeburden is forced on the user. Furthermore, in a case where theprocessing is executed by combining a dispenser, a centrifuge, amagnetic force device, a temperature controller, a device for sealingthe reaction vessel, a light measurement device, and the like, there isa concern of the scale of the utilized devices increasing and of thework area expanding. In particular, in a case where a plurality ofsamples is handled, since it becomes necessary to separate and extract aplurality of target nucleic acids and for amplification to be torespectively performed, the labor thereof becomes even greater, andfurthermore, there is a concern of the work area also expanding further.

Specifically, in a case where reactions of the nucleic acids (DNA, RNA,and the like) to be amplified, and the like, are performed within aplurality of reaction vessels and these reactions are monitored byoptical measurements, the measurements are performed by successivelymoving a single measuring device to the respective reaction vessels by amanual method, or the measurements are performed by providing ameasuring device to each of the respective reaction vessels beforehand.

In the former case where a single measuring device is used, when themeasuring device is attempted to be manually moved to the apertures ofthe reaction vessels, there is a concern of subtle differences occurringin the measurement conditions for each reaction vessel as a result ofsubtle displacements or relative motions between the reaction vessel andthe measuring device.

In the latter case where a measuring device is provided to each of therespective reaction vessels, although the positioning accuracy becomeshigh, there is a concern of the device scale expanding, and of themanufacturing costs increasing. Furthermore, although it is preferableto seal the apertures of the reaction vessels at the time of temperaturecontrol and the measurements, it is time-consuming to perform sealing,or opening and closing, with respect to a plurality of reaction vesselsby a manual method with a lid, and in particular, there is a concern ofthe lid becoming adhered to the vessel apertures such that it becomesdifficult to easily open the lid, and of contamination occurring fromthe liquid attached to the inside of the lid dripping or splashing.Furthermore, there is a concern of providing a dedicated opening andclosing mechanism of the lid, complicating the device, and increasingthe manufacturing costs (Patent Document 2).

As a device that automatically performs measurements without providing ameasurement device for each of the respective reaction vessels, there isa device that, at the time temperature control of a microplate having aplurality of wells is performed by a thermal cycler, successive lightmeasurements of the respective wells is performed by moving a detectionmodule over the microplate (Patent Documents 3 and 4).

In this device, since the detection module itself is moved in a state inwhich it is supported by the thermal cycler, a load that accompanies theacceleration from the movement is imparted on the detection module,which has precision optical system elements and electronic circuits suchas a photomultiplier tube, thus becoming the cause of noise orbreakdowns of the measurement device, and furthermore, there is aconcern of the device lifetime being shortened.

Moreover, since the detection module is supported by the microplate, oris supported by a lid body sealing the respective wells of themicroplate and only moves in a horizontal direction, a fixed spacingbetween the respective wells and the measuring end of the measurementdevice is necessary. Therefore, since attenuation from the scattering oflight, and the leakage or entry of light with respect to the adjacentwells cannot be completely blocked and prevented, there is a concern ofa measurement with a high accuracy not being able to be performed.

Furthermore, since the detection module divides the light path using ahalf mirror at the time light from the vessels is received or isirradiated, there is a need to take a long light path length within themeasurement device, and it has a problem in that there is a concern ofthe device scale becoming large.

In a case where the detection module is one that moves such that itpasses the respective wells that are arranged on the microplate and thenumber of wells becomes large, there is a concern of the processing timebecoming long due to the movement distance being long, and in addition,there is also a concern of the problems of the measurement devicementioned above occurring.

Moreover, at the time an optical measurement is performed on a sealedreaction vessel, there is a concern of the lid which has transparency,or the optical system elements, becoming cloudy from condensation, andthe measurements becoming difficult.

Consequently, in order to perform nucleic acid amplification and thelike, as a precondition thereof, specialized researchers or techniciansbecome necessary, and this situation is preventing the generalization ofgenetic analysis and the expansion of clinical applications inhospitals, and the like.

Therefore, at the time of clinical use and the like, in order to preventcross-contamination and to reduce user labor, and to easily perform fromthe extraction, the amplification, and further, by means of ameasurement, the genetic analysis of nucleic acids, then the automationof steps from the extraction of the target compound, reactions such asamplification, up to the measurements, the miniaturization of thedevice, and the provision of an inexpensive, high-accuracy device areimportant.

PRIOR ART DOCUMENTS Patent Documents

[Patent Document 1] International Publication WO96/29602

[Patent Document 2] Japanese Unexamined Patent Publication No.2002-10777

[Patent Document 3] U.S. Pat. No. 7148043

[Patent Document 4] U.S. Pat. No. 7749736

SUMMARY Problems to be Solved by the Invention

Therefore, the present invention is one that has been achieved in orderto solve the problems mentioned above. A first object thereof is inproviding an optical measurement device for a reaction vessel, and amethod thereof that, with regard to the optical state within a reactionvessel with respect to nucleic acids and the like, makes rapid andefficient measurements with a high accuracy and reliability possible.

A second object is in providing an optical measurement device for areaction vessel, and a method thereof that, by simplifying theconstruction of the optical system and performing measurements using asmall number of measurement devices with respect to a plurality ofreaction vessels, prevents the expansion of the device scale andincreases in the complexity of the device construction, and can beinexpensively manufactured and utilized.

A third object is in providing an optical measurement device for areaction vessel, and a method thereof that, by consistently automatingin parallel the optical measurements with respect to the plurality ofreaction vessels, in which reactions such as amplification of nucleicacids are performed, and the associated processing therein, processingwith a high reliability can be performed by preventing with certaintycontaminations due to the entry of contaminants into the plurality ofreaction vessels from the exterior, or liquid leakage from the pluralityof reaction vessels, and the contamination of light.

Means for Solving the Problem

A first aspect of the invention is an optical measurement device for areaction vessel comprising: a vessel group in which two or more reactionvessels are arranged; a light guide stage having two or more linkingportions to which the front ends of one or two or more light guideportions, which have a flexibility, that are directly or indirectlylinkable with the respective reaction vessels and optically connect withthe interior of the linked reaction vessels, are provided; a connectingend arranging body that has an arranging surface that arranges andsupports along a predetermined path two or more connecting ends, towhich back ends of the light guide portions, in which the front endsthereof are provided to the linking portions, are provided, theconnecting ends are provided corresponding to the respective linkingportions; a measurement device provided approaching or making contactwith the arranging surface that has one or two or more measuring endsthat are successively optically connectable with the respectiveconnecting ends along the predetermined path, and in which light basedon an optical state within the reaction vessels is receivable by meansof optical connections between the connecting ends and the measuringends; and a light guide switching mechanism that relatively moves therespective connecting ends arranged on the connecting end arranging bodyand the respective measuring ends such that they are successivelyoptically connected.

It is preferable for the vessel group to have in addition to thereaction vessels, two or more liquid housing parts that house liquidssuch as samples, reagents, and the like. Furthermore, the vessel groupincludes a microplate in which wells representing a plurality of liquidhousing parts are arranged in a matrix form or a column (row) form, or acartridge form vessel in which wells representing a plurality of liquidhousing parts are arranged in a row form. In a case where amplificationof nucleic acids is performed, the vessel group is provided with two ormore liquid housing parts housing for example; a sample, a magneticparticle suspension in which magnetic particles that are able to capturethe nucleic acids or the fragments thereof, which represent theamplification subject, are suspended, a solution for separating andextracting used for the separation and the extraction of theamplification subject, and an amplification solution used in nucleicacid amplification.

Here, the “amplification solution” represents, in a case whereamplification is performed by the PCR method for example, a template DNAsolution which is the amplification subject, a primer solution, a DNApolymerase solution, a nucleotide solution, a reaction buffer solution,and the like. In a case where amplification is performed by the SPIAmethod, it represents a DNA/RNA chimera primer solution, a DNApolymerase solution, an RNaseH solution, and the like. Furthermore,generally, methods for performing real-time PCR using fluorescentreagents containing a fluorescent compound include the intercalationmethod, the hybridization method, and the LUX method. In the“intercalation method”, a fluorescent compound such as SYBR (registeredtrademark) GREEN I or ethidium bromide, enters into double-stranded DNAat the time of the elongation reaction, and is a method in which the DNAamount is measured by irradiating an excitation light and utilizing thefluorescent light-emitting characteristics. Therefore, at the veryleast, the fluorescent material and a quencher that suppresses the lightemission of the fluorescent material must be contained within theamplification solution. The “hybridization method” is a method thatdetects only a target PCR product by using a DNA probe labeled with afluorescent material in addition to a PCR primer. That is to say, as aresult of the DNA probe labeled by the fluorescent material hybridizingwith the target PCR product, the hybridized DNA (amount) thereof isdetected. The “LUX method” is one that utilizes a property in which thefluorescent light signal of the fluorescent compound labeling theoligonucleotide is affected by the shape (such as a sequence, asingle-strand, or a double-strand) of the oligonucleotide thereof. Inactual real-time PCR, a PCR primer (LUX primer) that is labeled with onetype of a fluorescent compound and a contrastingly unlabeled PCR primerare used to perform real-time PCR. The LUX primer thereof is labeledwith a fluorescent compound in the vicinity of the 3′-terminus, and isdesigned such that it takes a hairpin structure in the interval betweenthe 5′-terminus. When the LUX primer takes a hairpin structure, thequenching effect is resolved, and the fluorescent light signal becomesincreased. By measuring this signal increase, the amount of the PCRproduct can be measured.

Examples of the material of the vessels, which includes the reactionvessels, the lid, and the like, include resins such as polyethylene,polypropylene, polystyrene and acrylic, glass, metals, and metalcompounds. The size of the vessels is, in addition to several μL toseveral 100 μL of liquid being storable, a size in which the ends of thedispensing tips are insertable for example. In the case of a cylindricalshape, the diameter of the size of one vessel is several mm to several10 mm, and the depth is several mm to several 10 mm for example.

It is preferable for the interior of the reaction vessels to betemperature controllable by a temperature controller.

The “temperature controller” has a temperature source that is able tolower the temperature within the reaction vessels, which house theliquids that become subjected to temperature control, based on a signalfrom the exterior for example. The temperature source is one in which,for example, a Peltier element, a heater, a cooling device, and the likeis provided on a block-shaped member. In order to perform processingsuch as PCR, the temperature controller is preferably a thermal cyclerusing a Peltier element. That is to say, it is preferable fortemperature control to be achieved by providing a block for temperaturecontrol as a temperature source to the vessel group or on the stage, inwhich temperature is raised and lowered by means of a Peltier element,that makes contact with or is adjacent to a portion (a lower side wallsection for example) or the entirety of the reaction vessel.Furthermore, it is also possible to perform temperature control of anisothermal amplification by means of the LAMP method.

“Temperature control” represents, with respect to a liquid or a vesselthat becomes the subject thereof, the maintaining of one or two or moreset predetermined temperatures for set time periods, according to aspecified sequence, and the execution at a specified frequency. Theinstructions to the temperature controller are carried out by sending acorresponding signal based on a program.

The “predetermined temperature” is a target temperature that an object,such as a liquid that becomes the subject, is to reach. In a case wherenucleic acids, such as the DNA contained in a liquid, oroligonucleotides and the like, which represent fragments of nucleicacids, are amplified by the PCR method for example, the predeterminedtemperature that is set is a temperature cycle performed in the PCRmethod. That is to say, it represents temperatures that are respectivelynecessary for the denaturation, the annealing or the hybridization, andthe elongation of DNA of approximately 94° C., a temperature in theinterval from 50° C. to 60° C., and a temperature of approximately 72°C. for example. On the other hand, in the case of the SPIA method(trademark), it becomes set at a fixed temperature, such as 55° C. forexample.

Furthermore, the predetermined temperature includes a temperature fortransition acceleration that shortens the transition time and keeps thesingle cycle time within a predetermined cycle time as a result of, inthe case of a transition from a high-temperature predeterminedtemperature to a low-temperature predetermined temperature, performingcooling at a temperature for transition acceleration that is lower thanthese predetermined temperatures by means of the temperature controller,or, at the time of a transition from a low-temperature predeterminedtemperature to a high-temperature predetermined temperature, byperforming heating at a temperature for transition acceleration that iseven higher than these predetermined temperatures for example. The“predetermined time” is the time necessary for maintaining therespective temperatures, and although it depends on the type of theamplification method, the reagents and the amount of liquid used in thePCR method, and the shape, the material, the size, the thickness, andthe like, of the nozzles, a single cycle is, in total, from severalseconds to several 10 seconds for example, and the processing time forthe PCR method as a whole is of the order of approximately severalminutes to several 10 minutes for example. The transition time is alsoincluded in the predetermined time.

The “linking portion” is a member that is able to be releasably linkedwith the reaction vessel directly, or indirectly via the sealing lid andthe like. Provided to the linking portion is the end of a light guideportion that is able to guide the light based on the optical statewithin the reaction vessel, by optically connecting with the interior ofthe reaction vessel. Here, the “linking with the reaction vessel”represents approaching or joining with the aperture, the outer wall, orthe outer bottom portion of the reaction vessel or a mounted sealing lidor sleeve and the like. Furthermore, “approaching” represents, withoutmaking contact, an approach to an extent that optical connection of theinterval with the light guide portions is possible. Moreover, “joining”includes making contact, close contact, adhesion, fitting, and mounting,and at the very least represents making contact such that opticalconnection of the interval between the light guide portions is possible.As a result of this linking, the light guide portion provided to thelinking portion and the interior of the reaction vessel are opticallyconnected. An example of the linking portion is a plate-shaped sectionof the light guide stage, and the end of the light guide portion is ahole piercingly provided in the plate-shaped section thereof, atransparent section such as an optical fiber, or an optical systemelement such as a lens. Alternatively, for example, it is a member of acylindrical shape, and the like, provided such that it protrudes fromthe light guide stage, and the end of the light guide portion is acavity provided in the member of a cylindrical shape, and the like, atransparent section such as an optical fiber, or an optical systemelement such as a lens. An example of a flexible light guide portion isan optical fiber or an optical fiber bundle. In a case where fluorescentlight is measured, it has two or more light guide portions, and aportion thereof is for irradiation, and the others are used forreceiving light. A case where it is directly linked with the aperture ofthe reaction vessel represents a case in which the interior of thereaction vessel is sealed using mineral oil and the like, and in thiscase, it is preferable to form the linking portion such that it is ableto directly seal the reaction vessel. Furthermore, in a case where thelinking is performed outside of the aperture, there is a need for thereaction vessel or the linking section thereof to have transparency.

The “predetermined path” represents, as a result of the measuring endsand the connecting end arranging body relatively moving, a path on aplane surface or a curved surface whereby the measuring ends are able toscan all of the connecting ends arranged therealong. Furthermore, thepath that connects all of the connecting ends represents a single ormultiple line segments that do not intersect (including zigzag lines andclosed straight lines), curved lines (including spirals and closedcurved lines), or a path along a combination of these and the like.Preferably, the respective single or multiple paths are continuous, andpaths along straight line segments without cusps or corners, or smoothcurves that have a curvature that the measuring ends are able to follow,are preferable.

There is a case where the linking portions and the connecting endscorrespond one-to-one, a case where they correspond many-to-one, and acase where they correspond one-to-many. Here, midway, it is possible forthe light guide portions to be branched or joined, or a light guideportion bundle comprising a plurality of light guide portions to bebranched or joined.

It is preferable for the predetermined path to be determined based onthe number, the shape, the arrangement, or the size of the measuringends on the measuring device, such that smooth scanning is possible. Forexample, a predetermined path along a straight line in which, for themovement of the connecting ends with respect to the measuring ends,there are no sudden changes in direction, such as changes to an obtuseangle or a right angle direction with respect to the travelingdirection, is preferable.

The arrangement pattern of the linking portions is a matrix form, acolumn form, or a row form for example. The arrangement pattern of theconnecting ends is the same arrangement, or a similar arrangement thatdiffers only in size for example, or in a case where the arrangementpattern is different, examples include the case of a circular form,other closed curved forms, a single column form, or a matrix form havinga smaller number of columns or rows. The predetermined path isdetermined such that it passes through all of the arranged connectingends.

Furthermore, it is preferable for the arrangement of the connecting endsto be such that they are integrated with respect to the arrangement ofthe linking portions. “Integration” is preferably performed by means ofthe predetermined path (or the arrangement pattern of the connectingends) representing a smaller region area or a smaller spacing than theregion area that encloses the arrangement pattern of the linkingportions on the light guide stage or the spacing between adjacentlinking portions, and by making the total scanning distance short.Consequently, in a case where the speed is made the same, processingwithin a shorter time than a case where the linking portions directlyscan the measuring ends is possible.

The extent of integration is preferably an extent in which the relativemovement or scanning of the connecting end arranging body and themeasuring device is able to complete the receiving of the light from allof the reaction vessels to be measured within the stable lightreceivable time for example. Here, the “stable light receivable time”represents the time in which the optical state within the reactionvessels, for which the light is receivable, is stably maintained. In thecase of the intercalation method or the LUX method of real-time PCR, orthe TaqMan probe of the hybridization method for example, it correspondsto the time in which the elongation reaction of the respective cycles ofPCR is performed. In a case where a FRET probe is used in thehybridization method, it corresponds to the time in which annealing isperformed.

Consequently, it can be applied with respect to a light emitter with ashort stable light receivable time and the like, and the versatility ishigh.

If the time taken for a single cycle is made several 10 seconds orseveral minutes for example, the stable light receivable time becomesseveral seconds or 10 seconds. However, the fluorescent light detectionamount of the initial cycles of a PCR reaction is below the detectionlimit, and the later cycles of the PCR reaction become a plateau state,and in order to secure quantitativeness by a strict definition, it mustbe within a range of the amplification curve in which an exponential PCRamplification can be observed. The present invention is one in which thestable light receivable time utilizes the fact that the movement time ofthe measuring end between the reaction vessels can be used, and byperforming the relative movement necessary for receiving the light fromthe respective reaction vessels within the stable light receivable time,the receiving of the light from the plurality of reaction vessels can beperformed approximately in parallel by means of a single measuringdevice, or a sufficiently small number in comparison to the number ofreaction vessels, without using complicated optical system elements andwithout expanding the scale of the device.

The “optical state” represents a state such as light emissions, colors,color changes, or light variations. The light based on the optical staterepresents light from light emissions or light variations, or reflectedlight from light irradiated with respect to colors or color changes, ortransmitted light, scattered light and the like.

The “connecting ends and the measuring ends are successively opticallyconnected” represents that the connecting ends and the measuring endsare optically connected by becoming opposed at a close proximity. Sincethe amount of light received by the measuring device at the moment ofconnection corresponds to a maximum value, the measurement controlportion specifies the data to be measured by calculating the maximumvalue of the amount of light.

The “measuring device” is one that makes fluorescence andchemiluminescence measurements possible for example, and in the formercase, it has a filter for the irradiation of one or two or more types ofexcitation light and the receiving of fluorescent light having one ortwo or more types of wavelengths. It is preferable for these to beguided using an optical fiber.

The “measuring end” has, at the very least, an inlet for the light to bereceived provided to the measuring device, and in the case of afluorescence measurement, has an outlet for the light to be irradiated.These can be provided as separate measuring ends. The inlet and theoutlet are respectively optically connected to a light receiving portioncomprising a photoelectric element provided in the interior, and to anirradiation source. At that time, they can be respectively connected viathe light guide portion for receiving light and the light guide portionfor irradiation. Furthermore, the connecting end arranging body, themeasuring ends, and the measurement device are preferably provided atseparated locations such that they do not directly make contact or arenot in the vicinity of the reaction vessels or the stage for mounting,in which heating control or temperature control is performed.

The light measurement device for a reaction vessel, although notexplicitly specified, additionally has “a measurement control portion”.The “measurement control portion” controls the measuring device and alight guide switching mechanism, comprises a computer (CPU) built intothe light measurement device for a reaction vessel and a program thatdrives the computer, and achieves measurement control by transmitting asignal through a DA converter to the respective control portions thatdrive the transfer mechanisms for example.

A second aspect of the invention is an optical measurement device for areaction vessel in which, at the time light is received by themeasurement device, at the very least, a measurement device bodyexcluding the measuring ends is immovably provided with respect to thelight guide stage, which is provided with the reaction vessels and thelinking portions linked with the same.

Therefore, there is a case where the connecting end arranging body moveswith respect to the measuring ends, or the measuring ends move withrespect to the connecting end arranging body, and the measurement devicebody may be movably provided with respect to the reaction vessels or thelight guide stage until the light guide stage is linked with thereaction vessels. The former case represents a case where themeasurement device body is linked with the light guide stage or a casewhere it is linked with movements in a portion of the directions forexample. Furthermore, the latter case represents a case where themeasurement device body is linked with the reaction vessels, or is fixedto the stage together with the reaction vessels. The measuring ends, ina case where they are present, include light guide portions, which areon the exterior of the measurement device body, up to the measuringends.

A third aspect of the invention is an optical measurement device for areaction vessel comprising a stage transfer mechanism that relativelymoves the light guide stage with respect to the vessel group, such thatthe linking portions are simultaneously directly or indirectly linkedwith two or more of the reaction vessels.

In a case where the stage transfer mechanism makes the light guide stagerelatively movable in the vertical direction with respect to the vesselgroup, the sealing lids that are mounted such that they cover theapertures of the reaction vessels can be pressed or shaken. That is tosay, it is preferable for the measurement control portion, followingindirectly linking with the linking portions via the sealing lids suchthat it covers the apertures of the reaction vessels, to perform controlsuch that it presses or shakes the sealing lids. By pressing, thesealing of the reaction vessels can be performed with certainty. Inaddition, by shaking, the sealed state between the apertures of thereaction vessels and the sealing lids can be rapidly and easily removedand released. Therefore, a high processing efficiency and reliabilitycan be obtained.

In a case where the linking portions are not linked by joining, such asby directly or indirectly fitting with the apertures of the reactionvessels, but by linking with the reaction vessels by approaching thereaction vessels, it is possible to successively and smoothly repeat thelinking between the linking portions and the reaction vessels, and thereleasing thereof, by means of horizontal movements without performingrelative movements in the vertical direction.

Furthermore, the two or more linking portions provided to the lightguide stage are arranged on a linking portion arranging body that ismovable in the horizontal direction with respect to the light guidestage in a state in which they are directly or indirectly simultaneouslylinkable with two or more reaction vessels. Moreover, by moving thelinking portion arranging body with respect to the light guide stage, anoptical measurement device for a reaction vessel that, without movingthe light guide stage, makes more reaction vessels linkable than thenumber of reaction vessels that are simultaneously linkable by thelinking portion arranging body, can be provided. In this case, it ispreferable for the linking between the respective linking portions andthe reaction vessels to be performed within shielded regions that aremutually shielded, such as within two or more grooves or two or moreregions that are mutually separated by a partition wall, to which therespective linking portions are insertable, that extend in a horizontaldirection in which the linking portion arranging body thereof ismovable, and are also provided to the light guide stage for each of thelinking portions. Consequently, the contamination of light from otherreaction vessels can be prevented with certainty.

In this case, the linking portions can be easily and rapidly linked withthe reaction vessels by merely moving in the horizontal directionwithout movements of the light guide stage in the vertical direction.Therefore, by setting the speed of the linking portion arranging bodysuch that it includes the horizontal movements of the linking portionarranging body and can be performed within the stable light receivabletime, it becomes possible to receive light and perform measurementsessentially in parallel with respect to an even greater number ofreaction vessels by means of a single set of measurement devices.

A fourth aspect of the invention is an optical measurement device for areaction vessel wherein the measurement device has: a plurality of typesof specific wavelength measurement devices that are provided with one ortwo or more measuring ends that are optically connectable with therespective connecting ends, and are able to receive light of specificwavelengths or specific wavelength bands; and a measuring end aligningportion that aligns the plurality of measuring ends such that they areoptically connectable with the respective connecting ends along thepredetermined path.

Here, in a case where fluorescent light is measured, the measurementdevice or the respective specific wavelength measurement devices areprovided with an excitation light irradiation source that irradiates thecorresponding excitation light, and a light receiving portion. Themeasuring end is provided with an irradiation aperture that connectswith the irradiation source, and a light receiving aperture thatconnects with the light receiving portion on the same measuring or as aseparate measuring end. The measuring ends are provided with a cavity,an optical element such as a lens, or a light guide portion such as anoptical fiber for example.

The “aligning” is integrally or linkingly performed. “Integrally”represents arrangement such that the intervals between the measuringends are mutually fixed and do not have any degrees of freedom.“Linkingly” represents arrangement such that the intervals between themeasuring ends have degrees of freedom to some extent, such as in achain. There is a case where the “aligning” is such that the respectivemeasuring ends are lined up along the scanning direction of thepredetermined path, or a direction perpendicular to the scanningdirection. In the latter case, the predetermined path is such that aplurality of paths are lined up in parallel.

According to the present aspect of the invention, by using a pluralityof types of luminescent compounds, colored compounds, color changingcompounds, or light variation compounds, and performing amplificationprocessing in parallel under the same conditions on a plurality of typesof amplification subjects in a single reaction vessel, it is possible toperform multiplex PCR amplification or multiplex real-time PCR on aplurality of types of amplification subjects by using a primer labeledwith a plurality of types of luminescent compounds for example.

Since it is “light of a specific wavelength or a specific wavelengthband”, it represents, in terms of visible light, a range of wavelengthssuch as a red light, a yellow light, a green light, a blue light or aviolet light for example.

A fifth aspect of the invention is an optical measurement device for areaction vessel wherein the vessel group is provided with sealing lids,which have transparency, that are mounted on apertures of one or two ormore of the reaction vessels, and seal the reaction vessels.

Here, the “sealing lid” includes, in addition to those that areinflexible and a plate form or block form, those that are a film form ora membrane form and have a flexibility. The “mounting” includes fitting,threading, friction, adsorption, attachment, adhesion, and the like. Inthese cases, detachable mounting is preferable.

Furthermore, in a case where the respective linking portions of thelight guide stage are linked at the apertures of the respective reactionvessels, it is preferable to make the linking portions or the nozzlespressable or shakable with respect to the sealing lids covering theapertures of the reaction vessels.

It is preferable for the linking portions to be provided such that theydownwardly protrude from the light guide stage. In this case, thelinking portions, for example, have a shape such as a rod shape, acylinder shape, a cone shape, and the like, and the lower end portionsof the members are preferably able to make contact with the sealinglids.

The sealing lids singularly cover the apertures of one or two or morereaction vessels. The sealing lids are moved by being mounted to thenozzles mentioned below, and cover the apertures of the reaction vesselsby using the tip detaching mechanism for example. To accomplish this,one or two or more indentations for mounting that are mountable on theone or two or more nozzles are provided on the upper side of the sealinglids. The one or two or more linking portions can be linked with thereaction vessels by being inserted within these indentations (which arealso indentations for linking) by means of vertical direction movementsof the light guide stage.

It also is possible, without moving the sealing lids by means of thenozzles, to provide a dedicated sealing lid transport mechanism. As thesealing lid transport mechanism, the optical measurement device for areaction vessel has a transporting body that is movable with respect tothe vessel group, and one or two or more grippers arranged on thetransporting body according to the arrangement of the reaction vesselsthat, with respect to the cover plate that covers the apertures of therespective reaction vessels, and the sealing lids having mountingportions that protrude on the lower side of the cover plate excludingthe center portion, in which light is able to pass through, and to whichthe cover plate is mountable to the reaction vessel, grips the coverplate such that the mounting portions are exposed on the lower side in astate in which they are mountable to the reaction vessels for example.Furthermore, if the sealing lid transporting body is made to be linkedwith the light guide stage, the device construction is simplified, andthe expansion of the device scale can be prevented.

In this case, since it is not necessary to provide an indentation formounting the nozzles and the like on the upper side of the sealing lid,the linking portion can be easily linked by just horizontal directionmovement between the apertures of the reaction vessels on the sealinglid without vertical direction movements of the light guide stage. Inthis case, if the horizontal direction movement of the linking portioncan be performed within the stable light receivable time, it becomespossible to receive light and perform measurements essentially inparallel with respect to an even greater number of reaction vessels.

A sixth aspect of the invention is an optical measurement device for areaction vessel, wherein the light guide stage is provided with aheating portion that is able to heat the sealing lids.

The measurement control portion, after simultaneously mounting thesealing lids to the linking portions, and following control of the stagetransfer mechanism such that the optical linking portions aresimultaneously indirectly linked with the two or more reaction vessels,controls the heating portion such that the sealing lids are heated forexample. The “heating portion” has a heating function at a temperaturethat is set based on the magnitude of an applied electric current or byan ON/OFF control for example.

Here, the heating of the sealing lids by the heating portion isperformed for preventing condensation at the time of temperature controlof the reaction vessels sealed by the sealing lids.

A seventh aspect of the invention is an optical measurement device for areaction vessel provided with a temperature controller having atemperature source that is provided making contact with or approachingthe lower side wall sections of the reaction vessels, and a heatingportion provided such that it is able to make contact with or approachthe upper side wall sections of the reaction vessels positioned furtheron an upper side than the lower side wall sections of the reactionvessels, and that has a heat source that is able to heat the upper sidewall sections.

Here, the “lower side wall section” represents a wall section or aportion thereof including the bottom portion that encloses a volumesection, which is a portion (1% to 90% for example) of the entire volumeof the reaction vessel in which a predetermined liquid amount determinedbeforehand is housable. The lower side wall section represents thesection of the wall section in which the rated liquid amount of liquidis housable for example. In a case where the reaction vessels comprisewide-mouthed piping parts that are linked with the linking portions, anda narrow-mouthed piping part, it is provided on the narrow-mouthedpiping part for example. The “upper side wall section” represents,within the entire volume of the reaction vessel, a vessel sectionenclosing the remaining volume of the lower side vessel section, inwhich the rated liquid amount is housed, or a portion thereof. The“upper side wall section” is normally preferably provided on the upperside of the reaction vessels leaving a spacing with the lower side wallsection. The upper side wall section becomes closer to the aperture, thesealing lid, or the linking portion than the lower side wall section. Inthe case of the vessel comprising the wide-mouthed piping part and thenarrow-mouthed piping part, and in a case where the linking portion islinked by fitting with the wide-mouthed piping part, the upper side wallsection is provided on the wall section of the wide-mouthed piping partfor example. The upper side wall section is a section that correspondsto a band shape along the circumference of the vessel wall for example.

The measurement control portion, following controlling the stagetransfer mechanism such that the linking portions are simultaneouslydirectly or indirectly linked with the reaction vessels, controls theheating portion such that direct or indirect condensation on the linkingportions is prevented. “Indirectly linked” represents a case where thelinking portions are linked with the reaction vessels via the sealinglids, the outer walls of the reaction vessels, and the like. “Control ofthe heating portion” is performed according to the “temperature control”for preventing condensation. For example, the heating temperature iscontrolled such that it is set from several degrees (a temperatureexceeding the dew point of water vapor that is necessary for preventingcondensation) to several 10° C. (a temperature that is sufficientlylower than the melting point of the raw material of the reactionvessels), for example, 1° C. to 60° C., or preferably approximately 5°C., higher than the respective predetermined temperatures set by thetemperature control for example. In a case where the amplification is byPCR, heating is performed at a temperature that is several degreeshigher than 94° C., such as 100° C., and in an isothermal case, in acase where the predetermined temperature is approximately 55° C.,heating is performed at a temperature that is several degrees higherthan this for example, such as from approximately 60° C. to 70° C.

As a result of the heating portion performing heating directly withrespect to the reaction vessels and not the linking portions or thesealing portions, thermal effects toward the optical system elementsprovided to the linking portion, or the measuring ends near the linkingportions, are reduced or removed. Therefore, the degradation of opticalsystem elements such as prisms, optical fibers, various lenses, such asconcavoconvex lenses, ball lenses, aspheric lenses, drum lenses, andgraded index rod lenses, mirrors, waveguide tubes, and the like, isprevented, and the reliability of the image that can be obtained throughthe optical system elements can be increased. For the linking portions,by using various lenses, such as ball lenses and aspheric lenses, whichrepresent the optical system elements, the light that is generatedwithin the reaction vessels and is outgoing in the aperture direction isfocused with certainty, and can be guided by being incident to the lightguide portion, such as an optical fiber.

Here, the reaction vessels; the temperature controller, which has atemperature source provided such that it is making contact with orapproaching the lower side wall sections of the reaction vessels, andperforms temperature control of the interior of the reaction vessels;and the heating portion provided such that it is making contact with orapproaching the upper side wall section, which has a heat source that isable to heat the upper side wall section, configure the reaction vesselcontrol system.

In that case, the reaction vessel preferably comprises a wide-mouthedpiping part, and a narrow-mouthed piping part that is formed narrowerthan the wide-mouthed piping part and is provided on a lower side of thewide-mouthed piping part and communicated with the wide-mouthed pipingpart. The end of the linking portion is fittable to the wide-mouthedpiping part, liquids are housable in the narrow-mouthed piping part, andthe lower side wall section is provided to the narrow-mouthed pipingpart, and the upper side wall section to the wide-mouthed piping part.Furthermore, it is preferable for the contact surfaces between thelinking portion and the upper side wall section of the reaction vessel,which is heated by the heating portion, or the sealing lid which makescontact therewith, to be made as small as possible. Consequently, theeffects of the heating portion toward the optical system elements of thelinking portions can be reduced or removed.

An eighth aspect of the invention is an optical measurement device for areaction vessel, wherein the light guide stage is provided to a nozzlehead having a suction-discharge mechanism that performs suction anddischarge of gases, and one or two or more nozzles that detachably mountdispensing tips in which the suction and the discharge of liquids ispossible by means of the suction-discharge mechanism, and has a nozzlehead transfer mechanism that makes the nozzle head relatively movablebetween the vessel groups.

In this case, in addition to further providing a magnetic force partthat is able to apply or remove a magnetic force within the dispensingtips mounted on the nozzles or liquid housing parts provided in thevessel group, and which is able to adsorb the magnetic particles on aninner wall of the dispensing tips or the liquid housing parts, it ispreferable to provide an extraction control part that controls thesuction-discharge mechanism, the transfer mechanism, and the magneticforce part, and as the reaction solution, separates and extracts thesolution of the amplification subject from the sample and houses itwithin the liquid housing parts as a portion of the amplificationsolution.

Here, the “solution for separating and extracting” includes a dissolvingsolution that breaks down or dissolves the protein forming the cellwalls and the like contained in the sample and discharges the nucleicacids or the fragments thereof to the outside of the bacteria or thecell, a buffer solution that simplifies the capture of the nucleic acidsor the fragments thereof by the magnetic particles, and additionally, asolution that dissociates from the magnetic particles, the nucleic acidsor the fragments of nucleic acids captured by the magnetic particles. Inorder to perform the separation of the nucleic acids or the fragmentsthereof, it is preferable to repeat the suction and the discharge of themixed solution.

The “dispensing tip” comprises for example a thick diameter portion, anarrow diameter portion, and a transition portion that communicatesbetween the thick diameter portion and the narrow diameter portion. Thethick diameter portion has an aperture for mounting, into which thelower end of the nozzle is inserted and the nozzle is mounted, and thenarrow diameter portion has an end mouth portion in which liquids canflow in and flow out by means of the suction and discharge of gases bythe suction-discharge mechanism. The dispensing tip and the nozzle aremanufactured from organic substances such as resins of polypropylene,polystyrene, polyester, acrylic, and the like, and inorganic substancessuch as glass, ceramics, metals including stainless steel, metalcompounds, and semiconductors.

The “suction-discharge mechanism” is for example a mechanism formed by acylinder, a piston that slides within the cylinder, a nut portion joinedto the piston, a ball screw on which the nut portion is threaded, and amotor that rotatingly drives the ball screw in both forward and reversedirections.

In a case where two or more nozzles are used, by respectively arrangingtwo or more vessel groups so as to correspond to the respective nozzleswithin two or more exclusive regions corresponding to the respectivenozzles, in which a single nozzle enters and the other nozzles do notenter, and by setting the respective exclusive regions for eachdifferent sample, cross-contamination between samples can be preventedwith certainty.

The stage transfer mechanism at least partly utilizes the nozzle headtransfer mechanism. It is preferable for the nozzle transfer mechanism,which moves the nozzles themselves in the Z axis direction, to also atleast partly utilize the nozzle head transfer mechanism, and for thestage transfer mechanism and the nozzle transfer mechanism to beindependently movable with respect to the Z axis direction movement.

A ninth aspect of the invention is an optical measurement device for areaction vessel, wherein the nozzle is such that a sealing lid isretainable by mounting, and by detaching the sealing lid, the sealinglid is mountable on an aperture of the reaction vessel. The detaching ofthe sealing lid can be combined with the tip detaching mechanism thatdetaches from the nozzle the dispensing tip mounted on the nozzle. Inthis case, the “sealing lid” has a sealing portion that is mountable onthe aperture of the reaction vessel, and an indentation for linking thatis mountable on the nozzle. Furthermore, in the case of the linkingportion indirectly linking with the reaction vessel by being mounted onthe sealing lid, in a case where the outer diameter of the nozzle andthe outer diameter of the linking portion are different, it ispreferable for the indentation for linking of the sealing lid to befitted with the end of the linking portion in place of the nozzle, bemounted, and make the linking portion able to retain the sealing lid. Inthis case, for the detaching of the sealing lid from the linkingportion, it is preferable to provide a dedicated sealing lid detachingmechanism.

A tenth aspect of the invention is an optical measurement device for areaction vessel, wherein front ends of a light guide portion bundle,which comprise a plurality of light guide portions, are provided to therespective linking portions, back ends of a light guide portion bundleof a portion of the light guide portion bundle are provided to firstconnecting ends of the connecting end arranging body, a portion or allof the remainder of the light guide portion bundle is provided to secondconnecting ends of the connecting end arranging body, the predeterminedpath comprises a first path and a second path, and by means of movementof the connecting end arranging body, first measuring ends provided onthe measurement device respectively relatively move along a first pathcomprising the first connecting ends, and second measuring ends along asecond path comprising the second connecting ends.

An eleventh aspect of the invention is an optical measurement device fora reaction vessel, wherein the first measuring end optically connectswith an irradiation source of the measurement device, the secondmeasuring end connects with a light receiving portion of the measurementdevice, an end corresponding to the first connecting end and an endcorresponding to the second connecting end are arranged such that theyare mixed, and the first measuring end is connectable with the firstconnecting end, and the second measuring end is connectable with thesecond connecting end.

Here, it is preferable for the “mixing of the ends” to be an arrangementsuch that the ends of the two or more types of light guide portions arehomogenized and intertwined.

twelfth aspect of the invention is an optical measurement device for areaction vessel, wherein the vessel group comprises two or moreexclusive regions corresponding to nozzles of respective groups, whichcomprise one or two or more nozzles, in which nozzles of a single groupenter and nozzles of other groups do not enter, and the respectiveexclusive regions at the very least have at least one of the reactionvessels, one or two or more liquid housing parts that house reactionsolutions used in the reactions, and sealing lids that are transportableto the reaction vessels using the nozzles and are able to seal thereaction solutions housed in the reaction vessels, and the light guidestage is extendingly provided across all of the exclusive regions suchthat linking portions of the light guide stage are associated such thatlinking portions of a single group, which comprises one or two or morelinking portions, enter the respective exclusive regions and linkingportions of other groups do not enter.

In order to make “the nozzles of a single group enter and the nozzles ofthe other groups not enter” or “the linking portions of a single groupenter and the linking portions of the other groups not enter”, forexample, this is performed by providing an exclusive region control partthat controls the nozzle head transfer mechanism such that nozzles of asingle group enter the respective exclusive regions and the nozzles ofthe other groups do not enter, and performs control such that thelinking portions of a single group enter the respective exclusiveregions and the linking portions of the other groups do not enter.

A thirteenth aspect of the invention is an optical measurement devicefor a reaction vessel that is further provided with a set of traversablenozzles comprising one or two or more nozzles that are movable such thatthey traverse the respective exclusive regions, and are able to enterthe respective exclusive regions.

A fourteenth aspect of the invention is an optical measurement devicefor a reaction vessel, wherein inspection information that identifiessamples or shows managed sample information and testing content isvisibly displayed at the respective exclusive regions, and a digitalcamera, which obtains image data by imaging the content displayed at therespective exclusive regions, which includes the sample information andthe testing content, is provided to the traversable nozzles.

Here, the “sample information” represents the information necessary foridentifying or managing the sample, and examples of the information foridentifying the sample include the attributes of the sample, such as thepatient, the animal, the food, the soil, the polluted water, or thelike, from which the sample was collected, and includes the name, theage, the sex, and the ID number of the patient, the sales location ofthe food, and the collection location and the collection date and timeof the soil, or the physical properties of the collected sample,including the classification of the blood, the urine, the faeces, thebodily fluid, the cells, or the like, of the patient, the classificationof the food, the classification of the soil, or the classification ofthe polluted water for example. Examples of the information that managesthe samples include the collector and the collection date of the samplethereof, the contact person for the sample, and the inspection date ofthe sample thereof for example.

The “inspection information” represents information showing the contentof the inspection performed with respect to the sample, and can includeinspection items such as various genetic information (SNPs, basesequence determination for example), genetic testing, or other variousprotein information or the types of reagents utilized in the inspection,the production lot number of the reagents, the calibration curves forthe reagents, the type and the structure of the instruments for testing,or the type of biological material fixed to a carrier and the like, forexample. This information is displayed in a handwritten case, a printedcase, a case where it is a barcode, or a case where it is a QR(registered trademark) code (a matrix type two-dimensional code) forexample. The image data is analyzed, converted to analysis datacorresponding to the code data, and output.

A fifteenth aspect of the invention is an optical measurement device fora reaction vessel comprising: a nozzle head provided with asuction-discharge mechanism that performs suction and discharge ofgases, and one or two or more nozzles that detachably mount dispensingtips in which the suction and the discharge of liquids is possible bymeans of the suction-discharge mechanism; a vessel group having at thevery least one or two or more liquid housing parts that house reactionsolutions used for various reactions, a liquid housing part that housesa magnetic particle suspension in which magnetic particles that are ableto capture a target compound are suspended, a liquid housing part thathouses a sample, one or two or more liquid housing parts that house asolution for separating and extracting of the target compound, and twoor more reaction vessels; a nozzle head transfer mechanism that makes aninterval between the nozzle head and the vessel group relativelymovable; a magnetic force part that is able to adsorb the magneticparticles on an inner wall of dispensing tips mounted on the nozzles; alight guide stage provided to the nozzle head and having two or morelinking portions to which ends of one or two or more light guideportions, which have a flexibility, that are directly or indirectlylinkable with the respective reaction vessels and optically connect withthe interior of the linked reaction vessels, are provided; a connectingend arranging body having an arranging surface that arranges andsupports along a predetermined path two or more connecting ends, towhich back ends of the light guide portions, in which front ends thereofare provided to the linking portions, are provided, the connecting endsare provided corresponding to the respective linking portions; ameasurement device provided approaching or making contact with thearrangement surface, having one or two or more measuring ends that aresuccessively optically connectable with the respective connecting endsalong the predetermined path, that is able to receive light based on anoptical state within the reaction vessels by means of opticalconnections between the connecting ends and the measuring ends; and alight guide switching mechanism provided along the predetermined path ofthe connecting end arranging body that relatively move the respectiveconnecting ends and the respective measuring ends such that they becomesuccessively optically connected.

Here, the “reaction solution” is for example an amplification solutionused for nucleic acid amplification. Furthermore, the “target compound”represents nucleic acids or the fragments thereof, which is theamplification subject. It is preferable to provide a tip detachingmechanism that detaches the sealing lids or the dispensing tips from thenozzles. In the present device, it is preferable to provide a samplesupplying device having a dispensing function that supplies the samples,the reagents, the washing liquids, the buffers, and the like that arenecessary for the vessel group at a position separate to the stage ofthe optical measurement device for a reaction vessel, and to make thewhole stage, to which the supplied vessel group is built-in, beautomatically moved to the position of the stage of the opticalmeasurement device for a reaction vessel and to be made exchangeable.Consequently, processing, including preparation processing such as thedispensing processing or the supplying processing with respect to thevessel group, can be consistently performed.

It is possible to respectively combine the second aspect of theinvention through to the thirteenth aspect of the invention with thepresent aspect of the invention.

A sixteenth aspect of the invention is an optical measurement method fora reaction vessel comprising: moving a light guide stage having two ormore linking portions provided with ends of one or two or more lightguide portions, which have a flexibility, with respect to two or morereaction vessels that are arranged in a vessel group; simultaneouslydirectly or indirectly linking the reaction vessels and the linkingportions and optically connecting the interior of the linked reactionvessels and the light guide portions; performing temperature controlwithin the reaction vessels; guiding light from the reaction vessels toa connecting end arranging body having an arranging surface thatarranges and supports along a predetermined path two or more connectingends, to which back ends of the light guide portions, in which frontends thereof are provided to the linking portions, are provided, theconnecting ends are provided corresponding to the respective linkingportions; and optically connecting along the predetermined path the oneor two or more measuring ends provided to a measurement device, whichare provided approaching or in contact with the arranging surface, andthe respective connecting ends, by moving the connecting end arrangingbody, to thereby make the measurement device receive light based on anoptical state within the reaction vessels.

It is possible to respectively combine the second aspect of theinvention through to the thirteenth aspect of the invention with thepresent aspect of the invention.

A seventeenth aspect of the invention is an optical measurement methodfor a reaction vessel, wherein the measurement device has a plurality oftypes of specific wavelength measurement devices that are able toreceive light of specific wavelengths or specific wavelength bands, andthe respective specific wavelength measurement devices have at least onemeasuring end that is successively optically connectable with theconnecting ends along the predetermined path. The method comprisesaligning the plurality of measuring ends by a measuring end aligningportion, and successively optically connecting the measuring ends withthe connecting ends along the path, to thereby make the respectivespecific wavelength measurement devices receive the light of specificwavelengths or specific wavelength bands based on an optical statewithin the reaction vessels.

An eighteenth aspect of the invention is an optical measurement methodfor a reaction vessel comprising simultaneously mounting two or moresealing lids, which have transparency, that are arranged in the vesselgroup and are fittable with apertures of the reaction vessels, onreaction vessels, and then moving the light guide stage with respect tothe sealing lids of the reaction vessels.

A nineteenth aspect of the invention is an optical measurement methodfor a reaction vessel comprising pressing or shaking with respect to thesealing lids covering the apertures of the reaction vessels.

A twentieth aspect of the invention is an optical measurement method fora reaction vessel comprising heating the sealing lids sealing thereaction vessels through the light guide stage.

A twenty-first aspect of the invention is an optical measurement methodfor a reaction vessel comprising: directly or indirectly linkingapertures of the reaction vessels and the linking portions; and at thetime of performing temperature control within the reaction vessels,according to temperature control by a temperature controller, which hasa temperature source provided making contact with or approaching lowerside wall sections of the reaction vessels, heating upper side wallsections of the reaction vessels, which are positioned further on anupper side than the lower side wall sections, by means of a heat sourceof a heating portion, which is provided making contact with orapproaching the upper side wall sections, and thereby preventing director indirect condensation of the linking portions.

A twenty-second aspect of the invention is an optical measurement methodfor a reaction vessel comprising: detachably mounting dispensing tips onrespective nozzles, which are provided to a nozzle head and perform thesuction and the discharge of gases; separating a target compound byusing a magnetic force part, a nozzle head transfer mechanism thatrelatively moves an interval between the nozzle head and a vessel group,a magnetic particle suspension housed within a vessel group, in whichmagnetic particles that are able to capture a target compound aresuspended, a sample, and a solution for separating and extracting of atarget compound; introducing the separated target compound and areaction solution used for reactions to a plurality of reaction vesselsprovided to a vessel group; moving a light guide stage, which has two ormore linking portions that are provided to the nozzle head and in whichfront ends of one or two or more light guide portions are also provided,with respect to apertures of the reaction vessels by means of, at thevery least, the nozzle head transfer mechanism; directly or indirectlysimultaneously linking the apertures of the reaction vessels and thelinking portions, and optically connecting the interior of the reactionvessels and the light guide portions that are linked; performingtemperature control within the reaction vessels; guiding light from thereaction vessels to a connecting end arranging body having an arrangingsurface that arranges and supports along a predetermined path two ormore connecting ends, to which back ends of the light guide portions, inwhich front ends thereof are provided to the linking portions, areprovided, the connecting ends are provided corresponding to therespective linking portions; and successively optically connecting oneor two or more measuring ends that are provided to a measurement device,and provided approaching or making contact with the arranging surface,and the connecting ends, along the predetermined path by relativelymoving them, to thereby make the measurement device receive the lightbased on an optical state within the reaction vessels.

It is possible to respectively combine the second aspect of theinvention through to the fourteenth aspect of the invention with thepresent aspect of the invention.

Effects of the Invention

According to the first aspect of the invention, the fifteenth aspect ofthe invention, the sixteenth aspect of the invention, or thetwenty-second aspect of the invention, as a result of linking with theplurality of reaction vessels by means of the linking portions providedto the light guide stage and optically connecting with the interior ofthe reaction vessels, the optical state within the reaction vessels istransmitted via the plurality of reaction vessels, the light guidestage, and the light guide portion, to the connecting ends of thearranging surface of the connecting end arranging body, and theconnecting ends arranged along the predetermined path on the arrangingsurface of the connecting end arranging body and the measuring ends ofthe measuring device are successively optically connected. Therefore,compared to a case where the measuring ends are directly scanned withrespect to the apertures of the reaction vessels, then in addition topreventing the attenuation or the leakage of light from the scatteringof light at the interval between the measuring ends and the liquidsurface, the arrangement of the connecting ends is such that it can berearranged in order to perform the connection with the measuring endsrapidly and smoothly, and with certainty. Therefore, measurements with ahigh reliability, and more efficient and rapid measurements of theoptical state within the reaction vessels, can be performed.

Consequently, with consideration of the stable light receivable time,the structure of the measuring ends, and the like, then the arrangingregion of the connecting ends as a whole, or the distance betweenadjacent connecting ends can be achieved by integration that makes thearranging region or the adjacent distances of the linking portionssmaller, and, by the smoothing of the movement of the measuring ends asa result of the linearization or the expansion of the radius ofcurvature of the predetermined path in comparison to the arrangement ofthe linking portion.

Switching of the optical system is performed by means of the movementbetween the measuring ends and the connecting ends on the arrangingsurface along the predetermined path. Therefore, the structure of theoptical system can be simplified. Furthermore, by separating theconnecting ends, the measuring ends, and the measurement device from thereaction vessels or the light guide stage, in which temperature controlor heating control is performed, thermal effects on the optical systemelements are excluded, and processing with a high reliability can beperformed.

The movement of the connecting ends with respect to the measuring endsincludes continuous or intermittent movement. As a result of themeasurement by real-time PCR, an amplification curve is created, whichcan be utilized in various analyses, such as the determination of theinitial concentration of DNA.

Moreover, since the measurement of the plurality of reaction vessels canbe performed in parallel with a single measuring device by utilizing thestable light receivable time, the expansion of the scale of the deviceis suppressed by reducing the number of measuring devices, and themanufacturing costs can be reduced. Further, since it is possible tomeasure, by successively moving the interval between the measuring endsand the connecting ends through the shortest distance along thepredetermined path determined beforehand, the measurements can beperformed in parallel by a simple mechanism of only a transfermechanism.

In a case where the reactions and the measurements are performed bysealing the reaction vessels by directly or indirectly linking theapertures of the reaction vessels with the linking portions, automaticmeasurements with a high reliability in which cross-contaminations andthe contamination of light can be prevented with certainty can beperformed.

According to the second aspect of the invention, at the time of movementwith respect to the connecting ends arranged on the connecting endarranging body and the measuring ends, the measurement device isimmovable with respect to the reaction vessels and the light guide stagethat is linked with the same. Therefore, at the time of a measurement, aload from an inertia force due to acceleration accompanying themovement, and the like, is not placed on the optical system elements orthe electronic elements built into the measurement device body,displacements of the optical system elements and destruction of theelectronic elements are prevented, and an accurate measurement with ahigh reliability can be performed. In cases other than a measurement,the measurement device body is movable with respect to the reactionvessels and the like. Therefore, it is possible to transport themeasurement device close to the reaction vessels and perform ameasurement.

According to the third aspect of the invention, the fifteenth aspect ofthe invention, the sixteenth aspect of the invention, or thetwenty-second aspect of the invention, by providing a stage transfermechanism that moves the light guide stage, it is possible tosimultaneously directly or indirectly link the linking portions with thereaction vessels without human intervention. Therefore,cross-contamination is prevented, and processing can be efficientlyperformed.

According to the fourth aspect of the invention, or the seventeenthaspect of the invention, by using a plurality of types of luminescentcompounds, colored compounds, color changing compounds, or lightvariation compounds, within a single reaction vessel, then for examplein a case where amplification processing is performed in parallel underthe same conditions on a plurality of types of amplification subjects,it is possible to perform multiplex PCR amplification or multiplexreal-time PCR on a plurality of types of amplification subjects by usinga primer labeled with a plurality of types of luminescent compounds andthe like. At that time, by combining the switching of the receiving ofthe light of a plurality of types of specific wavelengths or specificwavelength bands from the plurality of types of luminescent compoundsand the like, with a mechanism utilizing the stable light receivabletime that is used at the time of movement between the plurality ofreaction vessels, it is not necessary to separately provide a speciallight switching mechanism, and the device mechanism can be simplifiedand manufacturing costs can be reduced. Furthermore, since therespective specific wavelength measuring devices each receive light of aspecific wavelength or a specific wavelength band, the effects of otherspecific wavelengths or specific wavelength bands are not received, andhigh-accuracy measurements can be performed. Moreover, since therespective specific wavelength measuring devices are each modularizedsuch that removal and addition can be performed, processing with a highversatility according to the processing aims can be performed.

According to the fifth aspect of the invention or the eighteenth aspectof the invention, by mounting the sealing lids arranged in the vesselgroup to the linking portions or the nozzles, it is possible to performmounting to the apertures of the reaction vessels by means of movementof the nozzle head and the like. Therefore the housed substances withinthe reaction vessels do not make direct contact with the linkingportions of the stage, and hence cross-contaminations can be effectivelyprevented. Furthermore, since it is not necessary to provide a dedicatedmechanism for mounting the sealing lids on the reaction vessels, thescale of the device is not expanded, and the manufacturing costs arereduced.

According to the third aspect of the invention, or the nineteenth aspectof the invention, the sealing of the reaction vessels can be performedwith certainty by controlling the sealing lids covering the apertures ofthe reaction vessels such that they are pressed. Furthermore, by shakingthe sealing lids, the sealed state between the apertures of the reactionvessels and the sealing lids can be rapidly and easily removed andreleased. Therefore, a high processing efficiency and reliability can beobtained.

According to the sixth aspect of the invention, or the twentieth aspectof the invention, by performing control such that the linking portionsare heated, condensation at the time of temperature control of thereaction vessels that are sealed by the sealing lids is prevented, andmeasurements via the sealing lids, which have transparency, can beperformed with certainty and a high accuracy.

According to the seventh aspect of the invention, or the twenty-firstaspect of the invention, by performing heating of the upper side wallsections of the reaction vessels according to the temperature control ofthe lower side wall sections of the reaction vessels, the direct orindirect condensation of the linking portions can be prevented. In thiscase, the linking portions and the sealing lids are not directly heated,and heating is performed at the upper side wall sections of the reactionvessels. Therefore, the effects of direct heating toward the opticalsystem elements provided to the linking portions can be reduced orremoved. Consequently, in addition to reducing or removing imagedistortions and the like due to the degradation or the change inproperties of the optical system elements, various optical systemelements can be provided to the linking portions. Therefore, precisemeasurements with a high versatility can be performed. Furthermore, itis not necessary to provide a heating portion directly above thevessels, and the structure directly above the vessels, and therefore thestructure of the device as a whole is simplified, and it is possible tofurther approach the linking portions possessing optical systemelements, to the vessels and to perform the optical measurements withcertainty. With respect to the lower side wall sections, according tothe heating of the upper side wall sections, temperature control isperformed such that the temperatures are guided to the respectivepredetermined temperatures set using a coolable Peltier element and thelike, and measurements with a high reliability can be performed.

According to the eighth aspect of the invention, the twelfth aspect ofthe invention, or the fifteenth aspect of the invention, as a result ofthe light guide stage being built into the nozzle head to which thenozzles are provided, a transfer mechanism (at the very least for the Xaxis and Y axis directions) between the reaction vessels of themeasuring device is not separately provided, and since it can becombined with the transfer mechanism of the nozzles, the expansion ofthe scale of the device can be prevented. Furthermore, since thetransfer to the reaction vessel of the sample solution, the reagentsolutions, and the reaction solutions, which are to be housed within thereaction vessels, and which represent the measurement subject, and thepreparation, can be performed using the functions of the nozzles, stepsfrom the processing to the measurement of the measurement subject can beconsistently, efficiently, and rapidly performed.

According to the ninth aspect of the invention, since the sealing lidsare transferred by being mounted on the nozzles, a new, dedicated lidtransfer mechanism is not provided, and the expansion of the devicescale can be prevented by using existing mechanisms. On the other hand,in a case where the sealing lids are transferred or retained by beingmounted on the linking portions, it is possible to make the diameters ofthe linking portions and the nozzles different. Therefore, it becomespossible to provide various optical system elements within the linkingportions that are not limited by the size of the nozzles, and processingwith a high versatility and with reliability can be performed.

According to the tenth aspect of the invention, by providing the frontend of a light guide portion bundle comprising a plurality of lightguide portions to the linking portions, dividing the light guide portionbundle into a plurality of bundles, and separating the back end of thelight guide portions to a plurality of connecting ends, and bysimultaneously connecting with a plurality of measuring ends having oneor a plurality of measurement devices, the receiving of light of aplurality of types of wavelengths or wavelength bands, or theirradiation of excitation light with respect to the reaction vessels andthe receiving of light can be simultaneously performed. Therefore,processing of multiple fluorescent lights can be performed.

According to the eleventh aspect of the invention, the first measuringends optically connect with the irradiation source of the measurementdevice, and the second measuring ends optically connect with the lightreceiving portion of the measurement device, and in addition, the endsof the light guide portions that are connectable with the irradiationsource and the light receiving portion are mixed. Therefore, at the timeof a measurement of fluorescent light, it is possible to irradiateexcitation light within the reaction vessel without unevenness, and tomeasure the strength corresponding to the amount of fluorescence withcertainty.

According to the thirteenth aspect of the invention, by providingtraversable nozzles that are movable such that they traverse therespective exclusive regions, and dispensing target compounds or samplesof the same nucleic acids and the like with respect to a plurality ofexclusive regions, the same target compounds or samples can be utilizedin reactions in which the conditions are changed. Furthermore, bycombining the movement of the traversable nozzles with the transfermechanism of the connecting end arranging body, the expansion of thedevice scale can be suppressed.

According to the fourteenth aspect of the invention, by displayinginformation at the respective exclusive regions, and together with themovement of the traversable nozzles, reading in the informationdisplayed at the respective exclusive regions with a camera, reactionand measurement processing with a high reliability can be performedwithout expanding the device scale.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is an overall block-diagram showing an optical measurement devicefor a reaction vessel according to a first embodiment of the presentinvention.

FIG. 2 is an overall perspective view showing the optical measurementdevice for a reaction vessel according to a first exemplary embodiment.

FIG. 3 is a plan view showing enlarged, a vessel group of the opticalmeasurement device for a reaction vessel shown in FIG. 2.

FIG. 4 is a front view and a side view showing enlarged, a nozzle headof the optical measurement device for a reaction vessel shown in FIG. 2.

FIG. 5 is a perspective view as viewed from the front side of the nozzlehead shown in FIG. 4.

FIG. 6 is a side view showing more specifically, the transfer mechanismsand the suction-discharge mechanism shown in FIG. 4.

FIG. 7 is a perspective view showing more specifically, thesuction-discharge mechanism 53 and the like shown in FIG. 4.

FIG. 8 is a perspective view as viewed from the rear side of the nozzlehead shown in FIG. 4.

FIG. 9 is a cross-sectional view showing a state in which the linkingportion shown in FIG. 4 is linked with a reaction vessel.

FIG. 10 is a drawing showing the specific wavelength measurement deviceshown in FIG. 4.

FIG. 11 is a perspective view showing a light guide stage, a connectingend arranging body, and a sealing lid transport mechanism of an opticalmeasurement device for a reaction vessel according to a second exemplaryembodiment.

FIG. 12 is an enlarged perspective view showing the light guide stageshown in FIG. 11 with a portion cut away.

FIG. 13 is an enlarged cross-sectional view of the linking portion shownin FIG. 12.

FIG. 14 is a cross-sectional view showing an example of the sealing lidshown in FIG. 11.

FIG. 15 is an enlarged perspective view of the sealing lid transportingbody shown in FIG. 11.

FIG. 16 is a cross-sectional view of the sealing lid transporting bodyshown in FIG. 15.

FIG. 17 is a perspective view as viewed from the lower side of thesealing lid transporting body shown in FIG. 15.

FIG. 18 is a schematic diagram showing an example of the positionalrelationship between the optical fiber ends provided on the linkingportions and the reaction vessel.

FIG. 19 is an overall block-diagram showing an optical measurementdevice for a reaction vessel according to a second embodiment of thepresent invention.

FIG. 20 is a side view showing more specifically, the transfer mechanismand the suction-discharge mechanism according to a first exemplaryembodiment of FIG. 19.

FIG. 21 is a cross-sectional view showing a state in which a linkingportion according to the first exemplary embodiment of FIG. 19 is linkedwith a reaction vessel.

FIG. 22 is a cross-sectional view showing a state in which a linkingportion according to a second exemplary embodiment of FIG. 19 is linkedwith a reaction vessel.

FIG. 23 is a cross-sectional view showing a state in which a linkingportion according to a third exemplary embodiment of FIG. 19 is linkedwith a reaction vessel.

DETAILED DESCRIPTION BEST MODE FOR CARRYING OUT THE INVENTION

Next, an embodiment of the present invention is described with referenceto the drawings. This embodiment is not to be interpreted as limitingthe present invention unless particularly specified. Furthermore, in theembodiments or in the exemplary embodiments, the same objects aredenoted by the same reference symbols, and the descriptions are omitted.

FIG. 1 is shows a block-diagram of an optical measurement device for areaction vessel 10 according to a first embodiment of the presentinvention.

The optical measurement device for a reaction vessel 10 broadly has: avessel group 20 in which a plurality (twelve in this example) ofreaction vessel groups 23 i (i=1, . . . ,12, omitted hereunder) arearranged; a nozzle head 50 that has a nozzle arranging portion 70 inwhich a plurality (twelve in this example) of nozzles 71 i thatdetachably mount dispensing tips are arranged, and a light guide stage32 that has a plurality (twelve in this example) of linking portions 31i provided with the ends of two or more light guide portions, which havea flexibility, that are directly or indirectly linkable with theapertures of the reaction vessels and optically connect to the linkedreaction vessel interior; a measurement device 40 that is provided fixedto the nozzle head 50; a nozzle head transfer mechanism 51 that makesthe nozzle head 50 movable in the X axis direction for example; atemperature controller 29 that performs predetermined temperaturecontrol with respect to the reaction vessel group 23 i of the vesselgroup; a CPU+program 60 composed of a CPU, a ROM, a RAM, various typesof external memory, communication functions such as a LAN, and a programstored in the ROM, and the like; and a control panel 13 having a displayportion such as a liquid crystal display, and an operation portion, suchas operation keys or a touch panel.

The nozzle head 50 has: a stage Z axis transfer mechanism 35 that makesthe light guide stage 32 movable in the Z axis direction with respect tothe vessel group 20 independent of the nozzle arrangement portion 70; anozzle Z axis transfer mechanism 75 that makes the nozzle arrangementportion 70 movable in the Z axis direction with respect to the vesselgroup 20 independent of the stage 30; a magnetic force part 57 that, bymeans of a magnet 571 provided on a narrow diameter portion 211 ia of adispensing tip 211 i detachably mounted on the nozzle 71 i such that itcan approach and separate, is able to apply and remove a magnetic fieldwith respect to the interior; a suction-discharge mechanism 53 thatmakes the suction and the discharge of liquids with respect to thedispensing tip 211 i mounted on the nozzle 71 i possible by performingthe suction and the discharge of gases with respect to the nozzle 71 i;and a punching mechanism 55 which is driven by the suction-dischargemechanism 53, for punching a film that covers the apertures of theliquid housing parts of the vessel group 20 to seal various liquids inadvance. The stage transfer mechanism corresponds to the nozzle headtransfer mechanism and the stage Z axis transfer mechanism.

The nozzle head 50 further has a connecting end arranging body 30 thatarranges and supports a plurality (twelve in this example) of connectingends 34 i, which are provided corresponding to the respective linkingportions 31 i and are provided with the back ends of optical fibers(bundle) 33 i, which represent light guide portions in which the frontends thereof are provided to the linking portions 31 i, such that, as anarranging surface, they are integrated along a predetermined path (alinear path along the Y axis direction in this example) provided on avertical plane at a narrower spacing than the spacing between thelinking portions 31 i. Furthermore, the connecting end arranging body 30is provided at a position that is separated from the light guide stage32 and the reaction vessel group 23 i.

The measurement device 40 is able to respectively receive the light ofspecific wavelengths or specific wavelength bands of six types offluorescent light, and also has six types of specific wavelengthmeasurement devices 40 j (j=1, . . . ,6, omitted hereunder) that areable to irradiate excitation light of six types of specific wavelengthsor specific wavelength bands that are irradiated for the emission oflight.

The respective specific wavelength measurement devices 40 j havemeasuring ends 44 j that are provided approaching or making contact withthe arrangement surface, and are successively connectable with therespective connecting ends 34 i along the predetermined path (a linearpath along the Y axis direction). Furthermore, the respective measuringends 44 j have two ends, namely a first measuring end 42 j and a secondmeasuring end 43 j arranged along the Y axis direction. The firstmeasuring ends 42 j optically connect with an irradiation sourceprovided to the specific wavelength measurement devices 40 j. The secondmeasuring ends 43 j optically connect with a photoelectric device, suchas a photomultiplier tube, provided to the specific wavelengthmeasurement devices 40 j.

Furthermore, the nozzle head 50 has an arranging body Y axis transfermechanism 41, which represents a light guide switching mechanism, thatmoves the connecting end arranging body 30 along the Y axis direction onthe nozzle head 50 such that the respective connecting ends 34 iarranged on the connecting end arranging body 30 and the respectivemeasuring ends 44 j are successively connected.

Moreover, the light guide stage 32 has a heater 37, which represents theheating portion, for preventing condensation of the ends of the linkingportions 31 i or the mounted sealing lids 251 i, which havetransparency, by heating.

The vessel group 20 comprises a plurality (twelve in this example) ofexclusive regions 20 i, in which one (in this example, one groupcorresponds to one) nozzle enters and the other nozzles do not enter,that correspond to the respective nozzles. The respective exclusiveregions 20 i have: a liquid housing part group 27 i comprising aplurality of housing parts in which reagent solutions and the like arehoused or are housable; a sealing lid housing part 25 i in which one ortwo or more sealing lids 251 i, which have transparency, that aredetachably mounted on the nozzles are housed or are housable; andhousing parts for tips and the like 21 i that house, a plurality ofdispensing tips 211 i that are detachably mounted on the nozzles, andthe samples, and the like. The liquid housing part group 27 i has, atthe very least, one or two or more liquid housing parts that house amagnetic particle suspension, and two or more liquid housing parts thathouse a solution for separating and extracting used for the separationand the extraction of nucleic acids or the fragments thereof. Ifnecessary, it further has two or more liquid housing parts that house asolution for amplification used for the amplification of nucleic acids,and a liquid housing part that houses a sealing liquid for sealing thesolution for amplification housed in a PCR tube 231 i, which representsthe reaction vessel, within the PCR tube 231 i.

It is preferable for the exclusive regions 20 i to display a barcode asthe sample information and the inspection information for identifyingthe exclusive regions 20 i. Furthermore, the nozzle head 50 is providedwith a single traversable nozzle 710 in which liquids are transportableor dispensable by traversing (moving in the Y axis direction) theexclusive regions 20 i, and suction and discharge is made to beperformed by a traversable nozzle suction-discharge mechanism 17 that isseparate from the suction-discharge mechanism 53. Consequently, thesolution of DNA and the like housed in a given exclusive region 20 i canbe dispensed or delivered to the other exclusive regions 20 k (k≠i) Itis preferable for this movement in the Y axis direction to be also usedby the arranging body Y axis transfer mechanism 41.

The CPU+program 60 has, at the very least: a nucleic acid processingcontrol portion 63 that performs instructions for a series of processes,such as extraction and amplification with respect to nucleic acids orthe fragments thereof, and sealing of the solution for amplification,with respect to the temperature controller 29, the nozzle head transfermechanism 51, the tip detaching mechanism 59, the suction-dischargemechanism 53, the magnetic force part 57, and the nozzle Z axis transfermechanism 75; and a measurement control portion 61 that, after thenozzle head transfer mechanism 51 and the stage Z axis transfermechanism 35 are controlled such that the linking portions 31 i aresimultaneously directly or indirectly linked with the apertures of theplurality (twelve in this example) of PCR tubes 231 i, instructs ameasurement by the measurement devices 40 j by controlling the arrangingbody Y axis transfer mechanism 41 such that the optical fibers (bundle)33 i, which represent the light guide portions of the linking portions31 i, and the first measuring ends 42 j and the second measuring ends 43j of the measuring ends 44 j of the measurement devices 40 j mentionedbelow are optically connected.

Furthermore, the nucleic acid processing control portion 63 has anextraction control part 65 and a sealing lid control part 67. Thenucleic acid processing control portion 63 has the extraction controlpart 65 that performs instructions with respect to the tip detachingmechanism 59, the suction-discharge mechanism 53, the magnetic forcepart 57, the nozzle Z axis transfer mechanism 75, the nozzle headtransfer mechanism 51, and the stage Z axis transfer mechanism 35, for aseries of processes with respect to the nucleic acids or the fragmentsthereof, and the sealing lid control part 67 that performs instructionswith respect to the stage Z axis transfer mechanism 35 and the nozzlehead transfer mechanism 51 for a sealing process by the sealing lids.

Hereunder, a more specific first embodiment of the optical measurementdevice for a reaction vessel 10 mentioned above according to anembodiment of the present invention, is described with reference to FIG.2 to FIG. 10. FIG. 2 is a see-through perspective view showing anexternal view of the optical measurement device for a reaction vessel 10according to the first embodiment of the present invention.

FIG. 2A is a drawing showing an external view of the optical measurementdevice for a reaction vessel 10, which has: an enclosure 11 with a sizeof 500 mm in depth (Y axis direction), 600 mm in width (X axisdirection), and 600 mm in height (Z axis direction) for example, inwhich the vessel group 20, the nozzle head 50, a nozzle head transfermechanism 51 described in FIG. 1, and a CPU+program 60 are housed in theinterior; a control panel 13 provided on the enclosure 11; and a drawer15 to which a stage is provided.

FIG. 2B is a perspective view that sees through the interior of theenclosure 11, wherein the stage, into which the vessel group 20 isbuilt-in, is able to be drawn out to the exterior by means of the drawer15, and further, the nozzle head 50 is movably provided in the X axisdirection with respect to the vessel group 20 by means of the nozzlehead transfer mechanism 51 of FIG. 1.

FIG. 2B is a drawing showing that the nozzle head 50 is largely providedwith: various transfer mechanisms 52 having an arranging body Y axistransfer mechanism 41, a stage Z axis transfer mechanism 35, and anozzle Z axis transfer mechanism 75; a traversable nozzlesuction-discharge mechanism 17; the measuring device 40; a connectingend arranging body 30; an optical fiber (bundle) 33 i; and the magneticforce part 57. The traversable nozzle suction-discharge mechanism 17 andthe traversable nozzles 710 are supported such that they are movable inthe Y axis direction by means of the arranging body Y axis transfermechanism 41 such that they traverse the exclusive regions 20 i.

FIG. 3 is a plan view showing enlarged, the vessel group 20 shown inFIG. 2. The vessel group 20 is one in which twelve exclusive regions 20i (i=1, . . . ,12), wherein the longitudinal direction thereof is alongthe X axis direction and housing parts are arranged in a single rowform, are arranged in parallel along the Y axis direction at a pitch of18 mm for example. The exclusive regions 20 i are separately providedwith a cartridge vessel for PCR amplification 201 i, a cartridge vesselfor nucleic acid extraction 202 i, and a cartridge vessel for housingtips 203 i. The prevention of cross-contaminations between the exclusiveregions 20 i is achieved by providing partition walls 2010, 2020, and2030 on the cartridge vessels 201 i, 202 i, and 203 i of the exclusiveregions 20 i on the edge of one side along the X axis direction.

The cartridge vessel for PCR amplification 2011 has: the PCR tubes 231i, which represent the reaction vessel that are detachably linked withthe twelve linking portions 31 i provided to the light guide stage 32,via a single sealing lid 251 i which has transparency; the liquidhousing parts 271 i which house a buffer solution necessary for the PCRreaction; the sealing lid housing parts 25 i which house the sealinglids 251 i; the housing part for tips and the like 21 i that house thetips for punching for punching the film covering the PCR tubes 231 i andthe liquid housing parts 271 i, and the dispensing tips 211 i, andbarcodes 81 i that display the sample information and the inspectioninformation relating to the cartridge vessels for PCR amplification 201i.

The cartridge vessels for nucleic acid extraction 202 i has: sevenliquid housing parts 272 i for example, that house various reagents fornucleic acid extraction; reaction vessels 232 i that house the extractednucleic acids; and barcodes 82 i that display various information, suchas the sample information and the inspection information, related to thecartridge vessel. The PCR tubes 231 i and the reaction vessels 232 i aretemperature controllable by means of the temperature controller 29.

The cartridge vessels for housing tips 203 i has: a tip for punchingthat is able to punch the film covering the cartridge vessel for nucleicacid extraction 202 i; two small-quantity dispensing tips that performthe dispensing of small quantities of liquids; housing parts for tipsand the like 21 i that house dispensing tips for separations that areable to perform separation by adsorbing magnetic particles on an innerwall by applying and removing a magnetic force from the exterior, and abarcode 83 i that displays various information relating to the cartridgevessel 203 i.

The capacity of the PCR tube 231 i, which represents the reactionvessel, is of the order of approximately 200 μL, and the capacity of theother reaction vessels, liquid housing parts, and tubes is of the orderof approximately 2 mL.

The PCR tube 231 i is used for the amplification of nucleic acids or thefragments thereof, and temperature control is performed by means of thetemperature controller 29 based on a predetermined amplification method,such as a thermal cycle (from 4° C. to 95° C.) for example. The PCR tube231 i is formed with two levels as shown in FIG. 9 for example, and hasa narrow-mouthed piping part 233 provided on the lower side in which thesolution for amplification 234 i is housed, and a wide-mouthed pipingpart 235 i provided on the upper side in which the sealing lid 251 i isfittable. The inner diameter of the wide-mouthed piping part 235 i is 8mm for example, and the inner diameter of the aperture of thenarrow-mouthed piping part 233 i is approximately 5 mm for example. Thereaction vessels 232 i housed in the reaction tube housing holes aretemperature controlled for incubation to a constant temperature of 55°C. for example.

The liquid housing part group 272 i houses the solutions for separatingand extracting as follows. A first liquid housing part houses 40 μL ofLysis 1, a second liquid housing part houses 200 μL of Lysis 2, a thirdliquid housing part houses 500 μL of a binding buffer solution, a fourthliquid housing part houses a magnetic particle suspension, a fifthliquid housing part houses 700 μL of a washing liquid 1, a sixth liquidhousing part houses 700 μL of a washing liquid 2, a seventh liquidhousing part houses 50 μL of distilled water as a dissociation liquid,and an eighth liquid housing part, which is slightly separated, houses1300 μL of isopropyl alcohol (isopropanol) used for the removal ofprotein and the like, as a portion of the solution for separating andextracting protein. The respective reagents and the like are prepackedas a result of the punchable film covering the respective aperturesthereof.

In addition, 1.2 mL of distilled water is housed in a separate distilledwater reservoir, and tubes that house suspensions of bacteria, cells,and the like, or samples such as whole blood, are separately preparedfor each of the respective exclusive regions 20 i.

FIG. 4 is a front view and a side view of the nozzle head 50 accordingto the first embodiment of the present invention, and FIG. 5 is aperspective view from the front side.

The nozzle head 50 is one having: a nozzle arranging portion 70 in whichtwelve nozzles 71 i are arranged; a tip detaching mechanism 59 that isable to detach dispensing tips 211 i mounted on the nozzles 71 i; asuction-discharge mechanism 53; a magnetic force part 57 having twelvemagnets 571 provided such that they are able to approach and separatewith respect to the dispensing tips 211 i; a light guide stage 32;twelve linking portions 31 i provided to the light guide stage 32; atransfer mechanism portion 52 having a nozzle Z axis transfer mechanism75 and a stage Z axis transfer mechanism 35; optical fibers (bundles) 33i representing flexible light guide portions that extend to the rearside from the linking portions 31 i; a connecting end arranging body 30;the arranging body Y axis transfer mechanism 41; a measuring device 40having a measuring end 44; a traversable nozzle 710 ; and asuction-discharge mechanism 17 thereof.

The nozzle arranging portion 70 is provided with a cylinder supportingmember 73 that supports twelve cylinders 531 i such that they arearranged along the Y axis direction at a predetermined pitch of 18 mmfor example. The nozzles 71 i are provided on the downward end of thecylinders 531 i such that they are communicated with the cylinders 531i.

The tip detaching mechanism 59 is provided with detaching shafts 593 onboth sides, and has a tip detaching member 591 that detaches the twelvedispensing tips 211 i from the nozzles 71 i by sliding in the verticaldirection.

As shown specifically in FIG. 6 and FIG. 7, the tip detaching member 591is interlocked with the lowering of two tip detaching shafts 593 anddetaches the dispensing tips 211 i from the nozzles 71 i. The tipdetaching shaft 593 is elastically supported by the cylinder supportmember 73 by means of a spring 600 wrapped around the outer peripherysuch that it is biased in the upward direction, and the upper endthereof is positioned above the upper ends of the cylinders 531 i butbelow the lower limit position of the vertical movement range of thenormal suction and discharge of a cylinder drive plate 536 mentionedbelow. The two tip detaching shafts 593 are pushed in the downwarddirection by means of the cylinder drive plate 536 exceeding thevertical movement range and being lowered near the upper end of thecylinder 531 i, thus lowering the tip detaching member 591. The tipdetaching member 591 has twelve holes having an inner diameter that islarger than the outer diameter of the nozzles 71 i but smaller than themounting portions 211 ic, which represents the largest outer diameter ofthe dispensing tips 211 i, arranged at the pitch mentioned above suchthat the nozzles 71 i pass therethrough.

As shown specifically in FIG. 6 and FIG. 7, the suction-dischargemechanism 53 has: the cylinders 531 i for performing suction anddischarge of gases with respect to the dispensing tips 211 i which arecommunicated with the nozzles 71 i and mounted on the nozzles 71 i, anda piston rod 532 that slides within the cylinders 531 i; a drive plate536 that drives the piston rod 532; a ball screw 533 that threads withthe drive plate 536; a nozzle Z axis movable body 535 that, in additionto axially supporting the ball screw 533, is integrally formed with thecylinder support member 73; and a motor 534 mounted on the nozzle Z axismovable body 535 that rotatingly drives the ball screw 533.

The magnetic force part 57 has a magnet 571 that is provided such thatit can approach and separate with respect to the narrow diameterportions 211 ia of the dispensing tips 211 i detachably mounted on thenozzles 71 i, and is able to apply and remove a magnetic field in theinterior of the dispensing tips 211 i.

As shown specifically in FIG. 6, the nozzle Z axis transfer mechanism 75has: a ball screw 752 that threads with the Z axis movable body 535 andvertically moves the Z axis movable body 535 along the Z direction; anozzle head substrate 753 that axially supports the ball screw 752, andin addition to axially supporting the magnet 571 on the lower sidethereof such that it is movable in the X axis direction, is itselfmovable in the X axis direction by means of the nozzle head transfermechanism 51 mentioned below; and a motor 751 provided on the upper sideof the nozzle head substrate 753 that rotatingly drives the ball screw752.

As shown specifically in FIG. 6, the light guide stage 32 comprises ahorizontal plate 32 a and a vertical plate 32 b, which are letter-Lshaped plates in cross-section, and is provided with twelvecylinder-shaped linking portions 31 i having front ends of opticalfibers (bundles) 33 i, which are directly or indirectly linkable withthe apertures of the PCR tubes 231 i and are optically connected withthe interior of the linked PCR tubes 231 i, protruding in the downwarddirection from the horizontal plate 32 a. Furthermore, a heater 37 thatheats the sealing lids 251 i mounted on the linking portions 31 i andprevents condensation, is built into the bases of the linking portions31 i. The temperature of the heater is set to approximately 105° C. forexample. Since the light guide stage 32 is supported by the nozzle headsubstrate 753 by means of the nozzle head stage Z axis transfermechanism 35 such that it is movable in the Z axis direction, it ismovable in the nozzle X axis direction and Z axis direction.

The stage Z axis transfer mechanism 35 has: a side plate 355 provided onthe nozzle head substrate 753; a mount driving band-shaped member 354that is supported by a timing belt 352 spanning between two sprockets353 arranged in the vertical direction axially supported by the sideplate 355, and vertically moves in the Z axis direction; and a motorattached to the rear side of the nozzle head substrate 753 thatrotatingly drives the sprockets 353.

As shown in FIG. 7, the traversable nozzle suction-discharge mechanism17 is provided with a tip detaching mechanism 592 on the lower side ofthe suction-discharge mechanism 17 and on the upper side of the nozzle710. Furthermore, the suction-discharge mechanism 17 is provided with adigital camera 19. The suction-discharge mechanism 17 is movablyprovided in the Y axis direction by being attached to a timing belt 171spanning between two sprockets 173 that are rotatingly driven by a motor172.

FIG. 8 represents two perspective views of the nozzle head according tothe first embodiment viewed from the rear side, which show theconnection starting position (FIG. 8A) and the connection finishingposition (FIG. 8B) at the time the respective connecting ends of theconnecting end arranging body 30 and the respective measuring ends aresuccessively optically connected.

There are provided a connecting end arranging body 30 in which theconnecting ends 34 i provided corresponding to the respective linkingportions 31 i to which the front ends of the optical fibers (bundles) 33i which pass through the horizontal plate 32 a of the light guide stage32,are provided, and provided with the back ends thereof, are arrangedon an arranging surface on a path along a straight line in the Y axisdirection, which represents a predetermined path, at a shorter spacingthan the spacing of the linking portions 31 i; and six measuring endsthat are provided in the vicinity of, or making contact with, thearranging surface, and are successively optically connectable with theconnecting ends 34 i along the straight line. There is also provided ameasuring device 40 in which, by means of optical connections betweenthe connecting ends and the measuring ends, the fluorescent light withinthe PCR tubes 231 i, which represents the optical state, is receivable,and excitation light is also able to be irradiated.

Furthermore, the light guide stage 32 has a cylinder-shaped body 311 i,which retains the optical fibers (bundles) 33 i extending to the rearside from the linking portions 31 i such that they pass through theinterior in order to prevent folding, protrudingly provided upward fromthe horizontal plate 32 a directly above the linking portions 31 i. Inthe same manner, the connecting end arranging body 30 is also providedwith a cylinder-shaped body 301 i, which retains the optical fibers(bundles) 33 i extending from the connecting ends 34 i such that theypass through the interior in order to prevent folding, on the connectingend 34 i side.

The arranging body Y axis transfer mechanism 41 that moves theconnecting end arranging body 30 in the Y axis direction has: arms 412and 413 provided to the connecting end arranging body 30; a joining body411 that joins the arms 412 and 413 and the timing belt; a guide rail414 that guides the Y axis movement of the joining body 411; and twosprockets spanned by the timing belt and arranged along the Y axisdirection.

The measuring device 40 is one that supports the measurement offluorescent light and comprises six types of specific wavelengthmeasuring devices 40 j that are linearly aligned along a straight linein the Y axis direction, which represents the predetermined path, suchthat they support the measurement of six types of fluorescent light, andthey are provided fixed on a substrate of the nozzle head 50, such asthe frame that encloses the transfer mechanism portion 52, or a memberthat supports the same. Therefore, depending on the mechanism providedto the transfer mechanism portion 52, the measuring device 40 does notmove.

The measuring device 40 is one in which the measuring ends of theplurality of types (six in this example) of specific wavelengthmeasuring devices 40 j (j=1, 2, 3, 4, 5, 6), and therefore, in thiscase, the specific wavelength measuring devices 40 j themselves arealigned in a single row form, and integrally fixed to a member joinedwith the nozzle head substrate 753 using fixtures 45 j. The specificwavelength measuring devices 40 j have: measuring ends 44 j arrangedalong a straight line path in the Y axis direction which represents thepredetermined path, such that they successively optically connect to theconnecting ends 34 i; light detectors 46 j in which an optical systemcomponent having an irradiation source that irradiates excitation lightto the PCR tubes 231 i and a light receiving portion that receives thefluorescent light generated in the PCR tubes 231 i are built-in; andcircuit boards 47 j. The measuring ends 44 j have first measuring ends42 j that optically connect with the irradiation source, and secondmeasuring ends 43 j that optically connect with the light receivingportion. Here, the light detectors 46 j and the circuit boards 47 jcorrespond to the measuring device main body.

The pitch between the respective connecting ends 34 i, assuming a pitchbetween the linking portions 31 i of 18 mm, is 9 mm, which is halfthereof. Then, the pitch between the measuring ends 44 j is 9 mm or lessfor example.

There is a case where the first measuring ends 42 j and the secondmeasuring ends 43 j of the measuring ends 44 j of the respectivespecific wavelength measuring devices 40 j are arranged aligned in alateral direction (Y axis direction) along the straight line of the Yaxis direction along the predetermined path, and a case where they arearranged aligned in a longitudinal direction (X axis direction). In theformer case, without stopping the emission of the excitation light, therespective measuring devices successively receive light at a timing forreceiving light determined based on the speed of the connecting endarranging body, the pitch between the connecting ends, the distancebetween the first measuring ends and the second measuring ends of themeasuring ends, and the pitch between the measuring ends.

On the other hand, in the latter case, as shown in FIG. 8, with respectto the connecting end, a first connecting end and a second connectingend are provided. The first connecting end connects only with the firstmeasuring ends 42 j, and the second measuring ends 43 j connect onlywith the second connecting end. The fixed path represents two paths.Furthermore, the optical fibers (bundles) 33 i have optical fibers(bundles) 331 i for receiving light that have the first connecting end,and optical fibers (bundles) 332 i for irradiation that have the secondconnecting end. In this case, compared to the former case, connectionwith the linking portions is performed by means of optical fibers inwhich the irradiation source and the light receiving portion arededicated, and therefore, the control is simple, and the reliability ishigh since optical fibers that are respectively suitable for irradiationand receiving light can be used.

The speed of the connecting end arranging body 30 with respect to themeasuring ends 44 j is determined with consideration of the stable lightreceivable time, the lifetime of the fluorescent light with respect toexcitation light irradiation, the number of connecting ends, the pitchbetween the connecting ends, and the like (the distance of thepredetermined path). In the case of a real-time PCR measurement, it iscontrolled such that it becomes 100 mm to 500 mm per second for example.In the present embodiment, since the movement is performed by slidingthe arranging surface with respect to the measuring ends 44, theincidence of optical noise to the measuring ends 44 can be prevented.Furthermore, the connecting end arranging body 30 moves with respect tothe measuring ends intermittently such that it momentarily stops at eachpitch advance between the connecting ends or between the measuring ends,or continuously.

FIG. 9A is a drawing showing a state in which the linking portion 31 i(here, i=1 for example) that protrudes on the lower side from thehorizontal plate 32 a of the light guide stage 32 is indirectly linkedwith the PCR tube 231 i via the sealing lid 251 i, which hastransparency, that is mounted on the aperture of the PCR tube 231 i inthe exclusive region 20 i, and the linking portion 31 i is insertedwithin the indentation of the sealing lid 251 i, and the end surfacethereof is adhered to the bottom surface of the indentation of thesealing lid 251 i. The PCR tube 231 i comprises a wide-mouthed pipingpart 235 i and a narrow-mouthed piping part 233 i that is communicatedwith the wide-mouthed piping part 235 i and is formed narrower than thewide-mouthed piping part 235 i. Furthermore, the narrow-mouthed pipingpart 233 i is dried beforehand, or a liquid state solution foramplification 234 i is housed beforehand. Here, the reagent forreal-time amplification represents 70 μL of a master mix (SYBR(registered trademark) Green Mix) consisting of enzymes, buffers,primers, and the like.

For the aperture of the wide-mouthed piping part 235 i, since thesealing lid 251 i that protrudes on the lower side of the sealing lid251 i, which has transparency, is mounted on the reaction vessel, it ismounted to the reaction vessel as a result of a tubular sealing portion252 i, which encloses the center portion of the sealing lid 251 i inwhich light passes through, being fitted. At the time the sealingportion 252 i is fitted, it is preferable for the diameter of theoptical fibers (bundle) 33 i, which represents the light guide portionthat passes through the interior of the linking portion 31 i, to be thesame or larger than the diameter of the aperture of the narrow-mouthedpiping part 233 i. Consequently, it becomes possible to receive thelight from the PCR tube 231 i with certainty. The narrow-mouthed pipingpart 233 i is housed within a block for temperature control that isheated or cooled by means of the temperature controller 29.

In this example, the optical fibers (bundle) 33 i comprise opticalfibers (bundle) for irradiation 332 i that are connectable with thesecond measuring end 43 j and optical fibers (bundle) for receivinglight 331 i that are connectable with the first measuring end 42 j.

FIG. 9B is a drawing showing an example in which the optical fibers(bundle) 33 i comprise an optical fiber bundle in which an optical fiberbundle comprising a plurality of optical fibers for receiving light thatare connectable with the second measuring end 43 j, and an optical fiberbundle comprising a plurality of optical fibers for irradiation that areconnectable with the first measuring end 42 j, are combined such thatthey become uniform.

It is preferable for a feeding device for samples and the like to beintegrated to the optical measurement device for a reaction vessel 10.The feeding device for samples and the like is a device for dispensingand supplying parent samples and the like with respect to the vesselgroup 20, and the stage in which is integrated the vessel group 20 towhich the parent samples and the like have been supplied, isautomatically moved to the optical measurement device for a reactionvessel and the like. The feeding device for samples and the like, forexample, has a parent vessel group that houses parent samples and thelike, a tip detaching mechanism, a suction-discharge mechanism, and asingle nozzle that, in addition to the suction and the discharge ofgases being performed by means of the mechanisms, is detachably mountedwith dispensing tips 211 i. Furthermore, it has a nozzle head providedwith a mechanism that moves along the Z axis direction with respect tothe parent vessel group and the housing part group for tips and the like21 of the vessel group 20, an X axis movable body provided with a Y axistransfer mechanism that moves the nozzle head in the Y axis directionwith respect to the parent vessel group and the like, an X axis transfermechanism that moves the X axis movable body along the X axis withrespect to the parent vessel group and the like, and the parent vesselgroup. It is preferable for the parent vessel group to have a parentsample housing part group arranged in a 12 row ×8 column matrix formthat houses the parent samples to be supplied to the housing part groupfor tips and the like 21 of the vessel group 20, a distilled water andwashing liquid group, and a reagent bottle group.

FIG. 10 is a drawing showing a light detector 461 of a single specificwavelength measurement device 401 belonging to the measurement device 40according to a first exemplary embodiment of the present invention.

The specific wavelength measurement device 461 according to the presentexemplary embodiment, in addition to having an optical fiber 469 forexcitation light to be outgoing to the PCR tube 231 i and an opticalfiber 479 for light from the PCR tube 231 i to be incoming, has ameasuring end 441 provided on the lower ends of a first measuring end421 of the optical fiber 469 and a second measuring end 431 of theoptical fiber 479, an irradiation portion 462 that has a LED 467 thatirradiates excitation light through the optical fiber 469 and a filter468, and a light receiving portion that has an optical fiber 479, a drumlens 478, a filter 477, and a photodiode 472. This example shows a casewhere the first measuring end 421 and the second measuring end 431 areprovided along a direction (X axis direction) perpendicular to thestraight line of the Y axis direction, which represents thepredetermined path.

Next, a series of processing operations that perform real-time PCR ofthe nucleic acids of a sample containing bacteria using the opticalmeasurement device for a reaction vessel 10 according to the embodimentis described. Step S1 to step S13 below correspond to separation andextraction processing.

In step S1, the drawer 15 of the optical measurement device for areaction vessel 10 shown in FIG. 2 is opened, the vessel group 20 ispulled out, and by utilizing a feeding device for samples and the like,which is provided separately from the vessel group 20, the samples whichare subject to testing, various washing liquids, and various reagents,are supplied beforehand, and furthermore, a liquid housing part in whichreagents and the like are prepacked is mounted.

In step S2, following returning of the vessel group 20 and closing ofthe drawer 15, the start of the separation and extraction andamplification processing is instructed by means of the operation of thetouch panel of the control panel 13 for example.

In step S3, the extraction control part 65 provided to the nucleic acidprocessing controller 63 of the CPU+program 60 of the opticalmeasurement device for a reaction vessel 10 instructs the nozzle headtransfer mechanism 51 and moves the nozzle head 50 in the X axisdirection, positions the tip for punching mounted to the nozzle 71 iabove the first liquid housing part of the liquid housing part group 27i of the vessel group, and punches the film covering the aperture of theliquid housing part by lowering the nozzle by means of the nozzle Z axistransfer mechanism 75, and in the same manner, the other liquid housingparts of the liquid housing part group 27 i and the reaction vesselgroup 23 i are successively punched by moving the nozzle head 50 in theX axis direction.

In step S4, the nozzle head 50 is again moved in the X axis directionand moved to the housing part for tips and the like 21 i, and thenozzles 71 i are lowered by means of the nozzle Z axis transfermechanism 75, and the dispensing tips 211 i are mounted. Next, afterbeing raised by the nozzle Z axis transfer mechanism 75, the dispensingtips 211 i are moved along the X axis by means of the nozzle headtransfer mechanism 51, and advanced to the eighth liquid housing part ofthe liquid housing part group 27 i. Then a predetermined amount ofisopropanol is aspirated from the liquid housing part, and they areagain moved along the X axis direction, and predetermined amounts arerespectively dispensed into the solution components (NaCl, SDSsolutions) housed in the third liquid housing part and the fifth liquidhousing part, and the distilled water housed in the sixth liquid housingpart, so that 500 μL of a binding buffer solution (NaCl, SDS,isopropanol), 700 μL of a washing liquid 1 (NaCl, SDS, isopropanol), and700 μL of a washing liquid 2 (water 50%, isopropanol 50%) arerespectively prepared as solutions for separating and extracting withinthe third, the fifth, and the sixth liquid housing parts.

In step S5, following movement to, among the housing parts for tips andthe like 21 i, the sample tube in which the sample is separately housed,the narrow diameter portion 211 ia of the dispensing tip 211 i isloweringly inserted using the nozzle Z axis transfer mechanism 75, and,with respect to the suspension of the sample housed in the sample tube,following suspension of the sample within the liquid by repeating thesuction and the discharge by raising and lowering the drive plate 536 ofthe suction-discharge mechanism 53, the sample suspension is aspiratedwithin the dispensing tip 211 i. The sample suspension is moved alongthe X axis by means of the nozzle head transfer mechanism 51 to thefirst liquid housing part of the liquid housing part group 27 i housingthe Lysis 1 (enzyme) representing the solution for separating andextracting, and the narrow diameter portion 211 ia of the dispensing tip211 i is inserted through the hole in the punched film, and the suctionand the discharge is repeated in order to stir the sample suspension andthe Lysis 1.

In step S6, the entire amount of the stirred liquid is aspirated by thedispensing tip 211 i, and incubation is performed by housing it in thereaction vessel 232 i comprising the reaction tubes retained in thehousing holes, that is set to 55° C. by means of the constanttemperature controller. Consequently, the protein contained in thesample is broken down and made a low molecular weight. After apredetermined time has elapsed, the reaction mixture is left in thereaction tube, the dispensing tip 211 i is moved to the second liquidhousing part of the liquid housing part group 27 i by means of thenozzle head transfer mechanism 51, and the entire amount of the liquidhoused within the second liquid housing part is aspirated by using thenozzle Z axis transfer mechanism 75 and the suction-discharge mechanism53, and it is transferred using the dispensing tip 211 i by means of thenozzle head transfer mechanism 51, and the reaction solution isdischarged within the third liquid housing part by penetrating the holein the film and inserting the narrow diameter portion.

In step S7, the binding buffer solution housed within the third liquidhousing part, which represents a separation and extraction solution, andthe reaction solution are stirred, the solubilized protein is furtherdehydrated, and the nucleic acids or the fragments thereof are dispersedwithin the solution.

In step S8, using the dispensing tip 211 i, the narrow diameter portionthereof is inserted into the third liquid housing part by passingthrough the hole in the film, the entire amount is aspirated and thedispensing tip 211 i is raised by means of the nozzle Z axis transfermechanism 75, and the reaction solution is transferred to the fourthliquid housing part, and the magnetic particle suspension housed withinthe fourth liquid housing part is stirred with the reaction solution. Acation structure in which Na+ ions bind to the hydroxyl groups formed onthe surface of the magnetic particles contained within the magneticparticle suspension is formed. Consequently, the negatively charged DNAis captured by the magnetic particles.

In step S9, the magnetic particles are adsorbed on the inner wall of thenarrow diameter portion 211 ia of the dispensing tip 211 i byapproaching the magnet 571 of the magnetic force part 57 to the narrowdiameter portion 211 ia of the dispensing tip 211 i. In a state in whichthe magnetic particles are adsorbed on the inner wall of the narrowdiameter portion 211 ia of the dispensing tip 211 i, the dispensing tip211 i is raised by means of the nozzle Z axis transfer mechanism 75 andmoved from the fourth liquid housing part to the fifth liquid housingpart using the nozzle head transfer mechanism 51, and the narrowdiameter portion 211 ia is inserted by passing through the hole in thefilm.

In a state in which the magnetic force within the narrow diameterportion 211 ia is removed by separating the magnet 571 of the magneticforce part 57 from the narrow diameter portion 211 ia of the dispensingtip 211 i, by repeating the suction and the discharge of the washingliquid 1 (NaCl, SDS, isopropanol) housed in the fifth liquid housingpart, the magnetic particles are released from the inner wall, and theprotein is washed by stirring within the washing liquid 1. Thereafter,in a state in which the magnetic particles are adsorbed on the innerwall of the narrow diameter portion 211 ia as a result of approachingthe magnet 571 of the magnetic force part 57 to the narrow diameterportion 211 ia of the narrow diameter portion 211 ia again, thedispensing tip 211 i is, by means of the nozzle Z axis transfermechanism 75, moved from the fifth liquid housing part to the sixthliquid housing part by means of the nozzle head transfer mechanism 51.

In step S10, the narrow diameter portion 211 ia of the dispensing tip211 i is inserted by passing through the hole in the film using thenozzle Z axis transfer mechanism 75. By repeating the suction and thedischarge of the washing liquid 2 (isopropanol) housed in the sixthliquid housing part in a state in which the magnetic force within thenarrow diameter portion 211 ia is removed by separating the magnet 571of the magnetic force part 57 from the narrow diameter portion 211 ia ofthe dispensing tip 211 i, the magnetic particles are stirred within theliquid, the NaCl and the SDS is removed, and the protein is washed.Thereafter, in a state in which the magnetic particles are adsorbed onthe inner wall of the narrow diameter portion 211 ia by approaching themagnet 571 of the magnetic force part 57 to the narrow diameter portion211 ia of the dispensing tip 211 i again, the dispensing tip 211 i is,following raising by means of the nozzle Z axis transfer mechanism 75,moved from the sixth liquid housing part to the seventh liquid housingpart in which the distilled water is housed, by means of the nozzle headtransfer mechanism 51.

In step S11, the narrow diameter portion 211 ia of the dispensing tip211 i is lowered through the hole by means of the nozzle Z axis transfermechanism 75, and by repeating the suction and the discharge of thedistilled water at a slow flow rate in a state where the magnetic forceis applied within the narrow diameter portion 211 ia of the dispensingtip 211 i, the washing liquid 2 (isopropanol) is substituted by waterand is removed. Thereafter, by stirring the magnetic particles byrepeating the suction and the discharge within the distilled water whichrepresents the dissociation liquid, in a state in which the magnet 571of the magnetic force part 57 is separated from the narrow diameterportion 211 ia of the dispensing tip 211 i and the magnetic force isremoved, the nucleic acids or the fragments thereof retained by themagnetic particles are dissociated (eluted) from the magnetic particlesinto the liquid. Thereafter, a magnetic field is applied within thenarrow diameter portion and the magnetic particles are adsorbed on theinner wall by approaching the magnet 571 to the narrow diameter portion211 ia of the dispensing tip 211 i, and the solution containing theextracted nucleic acids, and the like, is made to remain in the eighthliquid housing part. The dispensing tip 211 i is moved to the storagepart of the housing parts for tips and the like 21 i in which thedispensing tip 211 i was housed, by means of the nozzle head transfermechanism 51, and the dispensing tip 211 i to which magnetic particlesare adsorbed, is detached from the nozzle 71 i together with themagnetic particles and dropped into the storage part, using thedetaching member 591 of the tip detaching mechanism 59.

The following step S12 to step S15 corresponds to nucleic acidamplification and measurement processing.

In step S12, a new dispensing tip 211 i is mounted on the nozzle 71 i,the solution housed within the eighth liquid housing part, whichcontains nucleic acids and the like, is aspirated, and by transferringit to the PCR tube 231 i in which the solution for amplification 234 iis housed beforehand, and discharging it, it is introduced into thevessel. As a result of moving the nozzle head 50 by means of the nozzlehead transfer mechanism 51, the nozzle 71 i is moved above the sealinglid housing part 25 i of the vessel group 20, which houses the sealinglid 251 i. Mounting is performed by lowering using the nozzle Z axistransfer mechanism 75 and fitting the indentation for linking 258 i onthe upper side of the sealing lid 251 to the lower end of the nozzle 71i. After being raised by the nozzle Z axis transfer mechanism 75, thesealing lid 251 is positioned above the PCR tube 231 i using the nozzlehead transfer mechanism 51, and by lowering the sealing lid 234 i bymeans of the nozzle Z axis transfer mechanism 75, it is fitted with theaperture of the wide-mouthed piping part 235 i of the PCR tube 231 i,mountingly sealing it.

In step S13, the nozzle head transfer mechanism 51 is instructed bymeans of an instruction from the measurement control portion 61, and bymoving the nozzle head 50 along the X axis, the linking portion 31 i ofthe light guide stage 32 is positioned above the PCR tube 231 i, whichis mounted with the sealing lid 251 i. Then, by lowering the light guidestage 32 by means of the stage Z axis transfer mechanism 35, the linkingportion 31 i is inserted into the indentation of the sealing lid 251 i,and the lower end thereof is made to make contact with, or adhere to,the bottom surface of the indentation.

In step S14, due to an instruction by the nucleic acid processingcontroller 63, the temperature controller 29 instructs a temperaturecontrol cycle by real-time PCR, such as a cycle in which the PCR tube231 i is heated for five seconds at 96° C. and heated for 15 seconds at60° C., to be repeated forty nine times for example.

In step S15, when temperature control at each cycle by the nucleic acidprocessing controller 63 is started, the measurement control portion 61determines the start of elongation reaction processing at each cycle,and instructs the continuous or intermittent movement of the connectingend arranging body 30 with respect to the measuring ends 44 j of themeasuring device 40. For the movement speed thereof, it is moved at aspeed that is calculated based on the stable light receivable time, thefluorescence lifetime, and the number (twelve in this example) ofexclusive regions 20 i. Consequently, the receiving of light from alltwelve PCR tubes 231 i within the stable light receivable time becomescompleted.

In step S16, the measurement control portion 61 determines the moment ofeach optical connection between the optical fibers (bundles) 33 i of thelinking portions 31 i and the first measuring end and the secondmeasuring end of the measuring end 44, and instructs the receiving oflight to the measuring device 40 for example.

This measurement is executed with respect to cycles in which exponentialamplification is performed, and an amplification curve is obtained basedon the measurement, and various analyses are performed based on theamplification curve. At the time of the measurement, the measurementcontrol portion 61 heats the heater 37 built into the light guide stage32 and prevents the condensation on the sealing lid 251, and a clearmeasurement can be performed.

FIG. 11 is a perspective view of the front side of a nozzle head 500 ofan optical measurement device for a reaction vessel according to asecond exemplary embodiment of the present invention, and a perspectiveview showing a portion thereof cut away. FIG. 12 is a perspective viewshowing enlarged, the portion of FIG. 11 shown cut away.

As shown in FIG. 11, in this example, unlike the optical measurementdevice for a reaction vessel according to the first exemplaryembodiment, the PCR tubes 236 i, which represent the reaction vessel,have a vessel group in which three or more rows of twelve each arearranged.

There are no large differences with the first exemplary embodiment withrespect to the section of the nozzle head 500 related to the dispensingdevice, which includes the nozzles, and the section related to thetraversable nozzle, the transfer mechanisms of the nozzle head, and thearranging body transfer mechanism, and they are omitted from thedescriptions. The nozzle head 500 has: a light guide stage 320; twelvelinking portions 310 i provided on the light guide stage 320; opticalfibers (bundle) 33 i that extend from the linking portions 310 i on therear side; a connecting end arranging body 300; a measuring device 400having a measuring end comprising six types of specific wavelengthmeasurement devices that are aligningly mounted on the light guide stage320; and a sealing lid transport mechanism 125.

The light guide stage 320 according to the second exemplary embodimenthas a linking portion arranging body 322, in which two or more (twelvein this example) linking portions 310 i that are simultaneously linkablewith two or more (twelve in this example) reaction vessels 236 i arearranged, that is movable in the horizontal direction (the X axisdirection in this example) with respect to the light guide stage 320.Furthermore, by means of the movement of the linking portion arrangingbody 322, without moving the light guide stage 320, it is linkable withmore reaction vessels 236 i (three rows of reaction vessels with twelveper row in this example) than the number of reaction vessels (twelve inthis example) that are simultaneously linkable by the linking portionarranging body 322.

The light guide stage 320 has a horizontal plate 320 a, a vertical plate320 b, and a triangular-shaped support side plate 320 c. The horizontalplate 320 a of the light guide stage 320, according to the arrangementof the linking portions 310 i arranged on the linking portion arrangingbody 322, is etchingly provided with two or more, or twelve in thisexample, long holes 321 i that correspond to shielding regions.

The measurement device 400 is mounted fixed to the upper edge of thevertical plate 320 b of the light guide stage 322. Therefore, since thelight guide stage 320 is stationary at the time of receiving light, themeasurement device 400 is immovably provided with respect to thereaction vessel and the light guide stage 320.

The optical fibers (bundle) 33 i having the end of the linking portion310 i, separate midway into optical fibers for receiving light (bundle)331 i and optical fibers for irradiation (bundle) 332 i. The opticalfibers for receiving light (bundle) 331 i connect to a second connectingend 341 i, and the optical fibers for irradiation (bundle) 332 i connectto a first connecting end 342 i, and are arranged as two paths along theY axis direction on a downwardly facing horizontal plane, whichrepresents an arranging surface on the lower side of the connecting endarranging body 300. At that time, the spacing between adjacentconnecting ends on these respective paths is such that they areintegrated at approximately half or one-third of the spacing of thelinking portions for example. The first connecting ends 342 i aresuccessively connectable with the first measuring ends of themeasurement device 400, and the second connecting ends 341 i aresuccessively connectable with the second measuring ends.

As shown in FIG. 12 or FIG. 13, the horizontal plate 320 a of the lightguide stage 320 is laminatingly provided with a thermal insulation plate323 formed by a resin and the like, a heater 370 provided on the lowerside of the thermal insulation plate 323 for preventing condensation ofthe sealing lids 253 i by heating the sealing lids 253 i, and athermally conductive metallic plate 325 provided on the lower side ofthe heater 370. Reference symbol 238 is a housing hole that houses thereaction vessels 236 i and is piercingly provided in the cartridgevessel. Reference symbol 239 represents a liquid surface that iscontrolled at a fixed height within the reaction vessels 236 i.Reference symbol 291 is a temperature controller for PCR.

The long holes 321 i that are etchingly provided in the horizontal plate320 a reach the metallic plate 325. Holes 326 that are the same size asthe apertures for light transmission are piercingly provided above theapertures of the reaction vessels 236 i of the metallic plate 325 of thebottom of the long holes 321 i, and are optically communicated with thebottom of the long holes 321 i.

The linking portions 310 i provided on the linking portion arrangingbody and the front ends of the optical fibers (bundle) 33 i provided inthe interior are, as a result of approaching the sealing lids 253 i,linked with the reaction vessels 236 i.

FIG. 14 is a drawing showing the various sealing lids 254 i to 257 iaccording to the second exemplary embodiment, that are mountable on thereaction vessel.

In FIG. 14A, the sealing lid 253 i has: a cover plate 25lia that coversthe aperture 236 ia of the reaction vessel 236 i; a central portion 253ic that is formed at the center of the cover plate 253 ic and thinnerthan the periphery, and has an increased light transmittance; and aclamp 253 ib comprising a double annular wall that is provided such thatit encloses the central portion 253 ic and protrudes on the lower side,that represents a mounting portion that is mountable to the outer edgeportion 236 ib of the aperture of the reaction vessel.

The sealing lid 254 i shown in FIG. 14B is formed thick in a convex lensform having a curved surface that expands from a central portion 254 ictoward the vessel exterior. Consequently, the light that is generatedwithin the reaction vessel is made to converge at the end of an opticalfiber, or the excitation light from the optical fiber is made toconverge at the liquid surface and the like, and the light can beefficiently collected.

The sealing lid 255 i shown in FIG. 14C is formed in a convex lens formhaving a curved surface that expands from a central portion 255 ictoward the vessel exterior, and consequently, the effects demonstratedin FIG. 14B are achieved.

The sealing lid 256 i shown in FIG. 14D is formed thick such that it hasa curved surface that expands from a central portion 256 ic toward thevessel exterior. Consequently, the effects demonstrated in FIG. 14B areachieved.

The sealing lid 257 i shown in FIG. 14E is formed such that it has acurved surface that expands from a central portion 257 ic toward thevessel exterior, and consequently, the effects demonstrated in FIG. 14Bare achieved.

FIG. 15 shows a sealing lid transporting body 125 according to thesecond exemplary embodiment.

The sealing lid transporting body 125 is one having: a prismaticsubstrate 128 that is movable in the X axis direction with respect tothe vessel group 20, which has at least three rows of reaction vessels236 i of twelve per row; one or two or more (twelve in this example)grippers 127 i arranged on the prismatic substrate 128 according to thearrangement of the reaction vessels that grip the cover plate such that,with respect to the sealing lid 253 i (to 256 i), the lower side isexposed in a state in which the mounting portion is mountable to thereaction vessel; and a bottom plate 126 that is mounted on the lowerside of the prismatic substrate 128.

As shown in the cross-sectional view of FIG. 16 and the perspective viewas viewed from the lower side of FIG. 17, the grippers 127 i have acavity 124 i that is cut out from the prismatic substrate 128 in anapproximate semicircular column shape such that most of the cover plate253 ia of the sealing lid 253 i is housable. Furthermore, the bottomplate 126 is provided a semicircular hole shaped notch portion 129 isuch that the clamp 253 ib, which represents the mounting portion of thesealing lid 253 i, is exposable on the lower side.

Next, the processing operation using the nozzle head 500 according tothe second exemplary embodiment is described.

Among the processes described in the first exemplary embodiment, theseparation and extraction process is omitted, and step S′12 to stepS′16, which correspond to nucleic amplification and measurementprocesses, are described.

In step S′12, a new dispensing tip 211 i is mounted on the nozzle 71 i,the solution containing nucleic acids and the like, which is housedwithin the eighth liquid housing part is aspirated, transported to thereaction vessel 236 i in which the solution for amplification 234 i ishoused beforehand, and discharged and introduced into the vessel. As aresult of moving the nozzle head 500 by means of the nozzle headtransfer mechanism 51, the sealing lids 253 i from the sealing lidhousing part in the sealing lid transporting body 125 in which twelvesealing lids 253 i are housed, are simultaneously housed in the cavity124 i of the grippers 127 i, and gripped.

Since the sealing lid transporting body 125 gripping the sealing lid 253i is linked with the light guide stage 320, then by using the stage Zaxis transfer mechanism 35 and moving it somewhat upwardly together withthe stage 320 and then moving it in the X axis direction, and bytransporting it to above the reaction vessels 236 i and lowering it, thetwelve sealing lids 253 i are sealed by mounting the clamps 253 ib,which are exposed on the lower side from the sealing lid transportingbody 125, to the PCR tubes 236 i. In the same manner, the rows of thetwenty four reaction vessels of the other two rows are successivelysealed by the sealing lids.

In step S′13, due to an instruction by the measurement control portion61, as a result of instructing the nozzle head transfer mechanism 51 andmoving the nozzle head 500 along the X axis, the light guide stage 320is moved such that it covers the thirty six reaction vessels of thethree rows, on which the sealing lids are mounted.

In step S′14, due to an instruction by the nucleic acid processingcontrol portion 63, the temperature controller 29 instructs atemperature control cycle by real-time PCR, such as a cycle in which thePCR tubes 231 i are heated for five seconds at 96° C. and heated forfifteen seconds at 60° C., to be repeated forty nine times for example.

In step S′15, when temperature control at each cycle is started by thenucleic acid processing control portion 63, the measurement controlportion 61 determines the start of the elongation reaction process ateach cycle, moves the linking portion arranging body 322 provided on thelight guide stage 320 over the light guide stage 320, indirectly linksthe respective linking portions 310 i that are inserted into the longholes 321 i provided on the light guide stage 320 via the reactionvessels and the sealing lids 253 i, and successively receives the lightfrom the reaction vessels while irradiating excitation light from themeasurement device to the interior of the reaction vessels. At the sametime, the continuous or intermittent movement of the connecting endarranging body 300 with respect to the respective measuring ends 44 j ofthe measurement device 400 is instructed. The movement speed thereof issuch that movement is performed at a speed that is calculated based onthe stable light receivable time, the fluorescence lifetime, the number(three rows of twelve reaction vessels per row in this example) ofreaction vessels of the exclusive regions 20 i that are measurable bythe light guide stage 320, and the like. Consequently, by moving thelinking portion arranging body 322 over the light guide stage 320 withinthe stable light receivable time, in this example, measurements can beperformed in parallel with respect to thirty six reaction vessels ofthree rows, with twelve per row.

In step S′16, the measurement control portion 61 determines the momentof the respective optical connections between the optical fibers(bundle) of the linking portions 310 i and the first measuring end andthe second measuring end of the measuring end 44, and instructs theirradiation of excitation light and the receiving of light to themeasurement device 400.

FIG. 18 is a drawing showing an example of the position of the opticalfiber front ends for receiving light and for irradiation provided to thelinking portion in a case where the linking portion is linked at alocation other than the aperture of the reaction vessel 236 i. FIG. 18Ais a drawing showing a case where the optical fibers (bundle) forreceiving light 331 i are in the vicinity of the outer bottom portion ofthe reaction vessel, and the optical fibers (bundle) for irradiation 332i are in the vicinity of the outer wall of the reaction vessel. FIG. 18Bis a drawing showing a case where the optical fibers (bundle) forreceiving light 331 i and the optical fibers (bundle) for irradiation332 i are in the vicinity of the outer wall of the reaction vessel. FIG.18C is a drawing showing a case where the optical fibers (bundle) forreceiving light 331 i and the optical fibers (bundle) for irradiation332 i are in the vicinity of the outer bottom portion of the reactionvessel. These are only examples, and cases where they are joined withthe reaction vessel by making contact, and the like, in place of beingin the vicinity are also possible.

FIG. 19 represents a block-diagram of an optical measurement device fora reaction vessel 110 according to a second embodiment of the presentinvention. Since the same reference symbols as the reference symbolsused in the first embodiment represent the same objects or similar (onlydiffering by size) objects, the descriptions thereof are omitted.

The optical measurement device for a reaction vessel 110 according tothe second embodiment differs from the optical measurement device for areaction vessel 10 according to the first embodiment in the aspect thatthe nozzle head 150 thereof has a light guide stage 132 that isdifferent from the light guide stage 32. The light guide stage 132according to the second embodiment differs from the light guide stage 32according to the first embodiment in the aspects that it has a plurality(twelve in this example) of linking portions 131 i in which the frontends of optical fibers, which represent two or more light guideportions, which have a flexibility, that optically connect with theinterior of the PCR tubes 231 i, and an optical element for collectinglight are provided in the interior, and the heat source of the heater137, which represents a heating portion for heating the reactionvessels, is provided not to the light guide stage 132, but to the vesselgroup 120 or the stage.

Further, it differs in the aspects that the sealing lids 251 i aretransported not by the nozzles 71 i but by fitting to the linkingportions 131 i, and are detached from the linking portions by means of adedicated sealing lid detaching mechanism 39. Therefore, the sealing lidcontrol portion 167, and therefore, the nucleic acid processing controlportion 163 and the CPU+program 160 differ from the device 10 accordingto the first embodiment.

The vessel group 120 is one in which twelve exclusive regions 120 i(i=1, . . . ,12), wherein the longitudinal direction thereof is alongthe X axis direction and housing parts are arranged in a single rowform, are arranged in the Y axis direction for example. The respectiveexclusive regions 120 i have: a reaction vessel group 23 i; a liquidhousing part group 27 i; a sealing lid housing part 25 i that housessealing lids 251 i, which have transparency, that are detachably mountedon the linking portions 131 i provided to the light guide stage 132; andhousing parts for tips and the like 21 i.

The reaction vessel 23 i, the temperature controller 29, and the heater137 correspond to the reaction vessel control system 90.

FIG. 20 is a cross-sectional view primarily showing, within the nozzlehead 150 according to a first exemplary embodiment of the secondembodiment, the transfer mechanism and the suction-discharge mechanism.

Here, since the diameter of the linking portion 131 i is thicker thanthe nozzle 71 i, the sealing lid 251 i to be mounted on the PCR tube istransported by the linking portion 131 i. Consequently, by utilizing thetransfer mechanism of the magnet 571 of the magnetic force part 57, asealing lid detaching mechanism 39 is provided that has a comb-shapeddetaching member 391 in which a notch portion, which has asemicircular-shaped arch that is approximately equivalent to thediameter of the twelve linking cylinders provided such that they canapproach and separate with respect to the linking portion 131 i, isarranged. In the present exemplary embodiment, since the detachment ofthe sealing lids can be performed by utilizing existing mechanisms, theexpansion of the device scale, and increases in complexity, can beprevented.

FIG. 21 is a drawing showing a reaction vessel control system 901according to the first exemplary embodiment of the second embodiment anda state in which, to the apertures of the reaction vessel group, towhich the PCR tubes 231 i representing a plurality (twelve in thisexample) of reaction vessels of the reaction vessel control system 901are provided, the linking portions 131 i (here, i=1 for example)protruding on the lower side from the horizontal plate 132 a of thelight guide stage 132 are indirectly linked with the PCR tubes 231 i viathe sealing lids 251 i, which have transparency, that are mounted on theapertures of the PCR tubes 231 i in the exclusive regions 120 i. As aresult of the linking portions 131 i fitting within the indentation forlinking 253 i of the sealing lids 251 i, they are linked with the PCRtubes 231 i.

As shown in FIG. 21, the linking portion 1311 is indirectly linked withthe PCR tube 231 i via the sealing lid 253, and has an approximatelycylinder-shaped linking cylinder 131 ai that is protrudingly providedfurther in the downward direction than the horizontal plate 132 a of thelight guide stage 132. Furthermore, a circular hole 131 bi having anaperture of a size corresponding to the liquid surface of the liquidthat is housed in the narrow-mouthed piping part is piercingly providedin the center portion of the bottom plate of the linking cylinder 131ai, and the periphery of the bottom plate is provided with a circularedge portion 131 di that is protrudingly provided below it.Consequently, the adhesion of the linking portion and the sealing lid isprevented. A spherical ball lens 381 i that has a diameter correspondingto the inner diameter of the linking cylinder is loosely inserted withinthe linking cylinder 131 ai and mounted on the circular hole 131 bi. Ata predetermined distance above the ball lens 381 i, an optical fiber 33i, in which the end is positioned and is covered by a resin-made ferrule131 ci that passes through the horizontal plate 132 a and reaches theexterior, is provided. The linking cylinder 131 ai, the circular hole131 bi, the ball lens 381 i, and the optical fiber 33 i bundle arearranged on the same axis in the interior of the linking cylinder 131ai.

As shown in FIG. 21, the reaction vessel control system 901 has: PCRtubes 231 i that represent reaction vessels, in which target solutionsof DNA having a target base sequence, and the like, are stored andreactions, such as amplification, are performed; a heater 137; and atemperature controller 291 i for PCR. The heater 137 is laminatinglyprovided with a heating block 137 c comprising an aluminum plate havinga high thermal conductivity, a sheet heater 137 a, and a heat insulator137 b. Twelve through holes 137 di that house and retain a plurality(twelve in this example) of PCR tubes 231 i are piercingly provided inthe same heater 137, and the wide-mouthed piping parts 235 i aresupported by the heating block 137 c.

The temperature controller 291 for PCR has: a block for temperaturecontrol 292 i that makes contact with, and is housable in, thenarrow-mouthed piping part 233 i of the PCR tube 231 i, which representsthe reaction vessel; a Peltier element 293 i; and a heat sink 294 i.

The narrow-mouthed piping part 233 i of the PCR tube 231 i has a lowerside wall section 233 ai of the section in which the block for PCR 292 iis making contact and is provided. Furthermore, it has an upper sidewall section 235 ai provided on the upper side leaving a spacing withthe lower side wall section 233 ai that corresponds to the wall sectionof the wide-mouthed piping part 235 i that makes contact with the blockfor heating 137 c of the heater.

According to the present exemplary embodiment, firstly, by means of aninstruction by the sealing lid control portion 167 (the CPU+program160), the nozzle head transfer mechanism 51 is instructed and followingmovement of the respective linking portions 131 i of the light guidestage 132 to the sealing lid housing parts 25 i, the stage Z axistransfer mechanism 35 is instructed and the sealing lids 251 i arefitted and mounted to the linking portions 131 i. Next, by fitting theapertures of the predetermined PCR tubes 231 i with the sealing lids 251i, the linking portions 131 i are simultaneously linked with the PCRtubes 231 i.

Next, according to the temperature control by the temperature controller29 as a result of an instruction by the measurement control portion 161,in the case of PCR, by controlling the heater 137 such that the upperside wall section 233 bi is heated at a fixed temperature (100° C. forexample) that is several degrees, or preferably approximately 5° C.,higher than the maximum predetermined temperature (94° C. for example),the sealing lid 251 i fitted to the wide-mouthed piping part 235 i ofthe PCR tube 231 i is heated, and condensation of the sealing lid can beprevented. At that time, the upper side wall section 235 ai is separatedby a fixed spacing from the lower side wall section 233 ai, in whichtemperature control is performed, and the upper side wall section 233ai, which has a smaller surface area than the lower side wall section,is heated by bringing the heat source into contact or into its vicinity.Consequently, the effect of heating the upper side walls section 235 aiis such that the lower surface of the sealing lid 251 i, which isprovided at a position near the upper side walls section 235 ai, isheated, and condensation can be prevented.

On the other hand, since the linking portion 131 i is only makingcontact with the upper side of the sealing lid 251 i via the circularedge portion 131 di, the effect of heating is not as much as withrespect to the sealing lid 251 i. In the same manner, the lower sidewall section 233 ai is temperature controlled to the predeterminedtemperature using a Peltier element having a heating and coolingfunction, and furthermore, measurements are simultaneously performed.After completion of the measurement, then by means of an instruction bythe sealing lid control portion 167, the linking portion 131 i is madeto approach using the detaching member 391, and then by upwardly movingthe light guide stage 132 by means of the stage Z axis transfermechanism 35, the sealing lid 251 i is detached from the linking portionand while remaining on the PCR tube 231 i, the linking portion is movedand the linking is released.

FIG. 22 is a drawing showing a second exemplary embodiment, andrepresents a linking portion 131 i in which, in place of the ball lens381 i, a drum lens 382 i having a lens diameter corresponding to theinner diameter of the linking cylinder 131 ai is loosely inserted withinthe linking cylinder 131 ai and mounted on the circular hole 131 bi, andis provided such that light is collected at the end of the optical fiber33 i.

FIG. 23 is a drawing showing a third exemplary embodiment, andrepresents a linking portion 131 i in which, in place of the ball lens381 i and the like, an aspheric surface lens 383 i having a lensdiameter corresponding to the inner diameter of the linking cylinder 131ai is loosely inserted within the linking cylinder 131 ai and mounted onthe circular hole 131 bi, and is provided such that light is collectedat the end of the optical fiber 33 i. Reference symbol 391 represents acomb-shaped detaching member of the sealing lid detaching mechanism 39,and shown is a state in which it is in the vicinity of, or makingcontact with, the linking portion 131 i. In this state, by raising thelinking portion 131 i, the sealing lid 251 i engages with the sealinglid detaching member 391 and is detached from the linking portion 131 i,but remains still mounted on the PCR tube 231 i. Furthermore, therespective lenses 381 i to 383 i may be made to be loosely mountedwithin the linking cylinder 131 ai by installing a tube-shaped framefrom the upper side.

The foregoing exemplary embodiments have been specifically described inorder to better understand the present invention, and they are in no waylimiting of other embodiments. Therefore, modifications are possiblewithin a scope that does not depart from the gist of the invention. Theconfigurations, shapes, materials, arrangements, and amounts of thenozzles, the dispensing tips, the punching tips, the vessel group, theexclusive regions thereof, the housing parts, the measuring ends, themeasurement devices, the specific wavelength measurement devices, thesuction-discharge mechanism, the transfer mechanism portion, themagnetic force part, the heating portion, the reaction vessels, thesealing lids, the light guide stage, the linking portions, the lightguide portions, the connecting ends, the connecting end arranging body,the linking portion arranging body, the nozzle head, the temperaturecontroller, the nozzle detaching mechanism, and the sealing liddetaching mechanism, and the like, and the utilized reagents and samplesare also in no way limited by the examples illustrated in the exemplaryembodiments. Furthermore, although the nozzles were made to move withrespect to the vessel group, it is possible to also move the vesselgroup with respect to the nozzles.

Furthermore, in the foregoing descriptions, although the amplificationsolution was sealed using a sealing lid for the sealing of the reactionvessel for PCR, it may be made such that, in its place or incombination, it is sealed using a sealing liquid, such as mineral oil.Furthermore, in place of punching by mounting a tip for punching on thenozzles, it is possible to use a punching pin that is driven by thesuction-discharge mechanism. Moreover, although a real-time PCRmeasurement was described in the foregoing descriptions, it is in no waylimited to this measurement, and it can be applied to other variousmeasurements in which temperature control is performed. In the foregoingdescriptions, although a case where the measurement device is providedto a dispensing device was described, it is not necessarily limited tothis. Although only an optical system using optical fibers wasdescribed, it is possible to also employ an optical system using a lenssystem in the interior of the measurement device.

Furthermore, the devices described in the respective exemplaryembodiments of the present invention, the components that form thesedevices, or the components that form these components, can beappropriately selected, and can be mutually combined by applyingappropriate modifications. The spatial representations within thepresent application, such as “above”, “below”, “interior”, “exterior”,“X axis”, “Y axis”, and “Z axis” are for illustration only, and are inno way limiting of the specific spatial directions or arrangements ofthe construction.

INDUSTRIAL APPLICABILITY

The present invention is related to fields in which the processing,testing, and analysis of nucleic acids, which primarily includes DNA,RNA, mRNA, rRNA, and tRNA for example, is required, and is related toindustrial fields, agricultural fields such as food, agriculturalproducts, and fishery processing, chemical fields, pharmaceuticalfields, health care fields such as hygiene, insurance, diseases, andgenetics, and scientific fields such as biochemistry or biology forexample. The present invention is, in particular, able to be used inprocessing and analysis that handles various nucleic acids, and thelike, such as PCR and real-time PCR.

BRIEF DESCRIPTION OF THE REFERENCE SYMBOLS

10, 110 Optical measurement device for reaction vessel

20, 120 Vessel group

20 i, 120 i (i=1, . . . ,12) Exclusive regions

211 i (i=1, . . . ,12) Dispensing tips

231 i, 236 i (i=1, . . . ,12) PCR tubes (reaction vessels)

30, 300 Connecting end arranging body

31 i, 131 i (i=1, . . . ,12) Linking portions

32, 320, 132 Light guide stage

33 i Optical fibers (light guide portions)

40, 400 Measurement device

40 j (j=1, . . . ,6) Specific wavelength measurement devices

44 Measuring end

50, 500, 150 Nozzle head

52 Transfer mechanism portion

53 Suction-discharge mechanism

59 Tip detaching mechanism

61, 161 Measurement control portion

70 Nozzle arranging portion

71 i (i=1, . . . ,12) Nozzle

What is claimed is:
 1. An optical measurement device, comprising: afirst optical fiber including opposing first and second ends; a secondoptical fiber including opposing third and fourth ends; a first reactionvessel comprising an opening to a first interior that is opticallyconnectable with the first end of the first optical fiber; a secondreaction vessel comprising an opening to a second interior that isoptically connectable with the third end of the second optical fiber; alight guide stage comprising a light guide plate coupled to the firstend of the first optical fiber and the third end of the second opticalfiber, wherein the light guide stage is driven in a vertical directionrelative to the openings of the first and second interiors by a stage Zaxis transfer mechanism to simultaneously optically connect: the firstend of the first optical fiber with the first interior of the firstreaction vessel, and the third end of the second optical fiber with thesecond interior of the second reaction vessel; a controller operablyconnected to the light guide stage; a first measurement device forreceiving emissions from the first and second reaction vessels, thefirst measurement device including a first photoelectric element and afirst inlet optically connected with the first photoelectric element;and a connecting end arranging body that is operatively connected to thecontroller and that arranges and supports the second end of the firstoptical fiber and the fourth end of the second optical fiber along afirst predetermined path, wherein the connecting end arranging body isdriven by the controller along the first predetermined path by anarranging body Y axis transfer mechanism between: a first measurementposition, in which the second end of the first optical fiber isoptically connected with the first photoelectric element via the firstinlet so that light based on an optical state within the first reactionvessel is transmittable from the first reaction vessel to the firstphotoelectric element, via at least the first optical fiber and thefirst inlet; and a second measurement position, in which the fourth endof the second optical fiber is optically connected with the firstphotoelectric element via the first inlet so that light based on anoptical state within the second reaction vessel is transmittable fromthe second reaction vessel to the first photoelectric element, via atleast the second optical fiber and the first inlet.
 2. The opticalmeasurement device of claim 1, further comprising: a second measurementdevice including a second photoelectric element and a second inletoptically connected with the second photoelectric element; wherein theconnecting end arranging body is driven by the controller along thefirst predetermined path by the arranging body Y axis transfer mechanismbetween: a third measurement position, in which the second end of thefirst optical fiber is optically connected with the second photoelectricelement via the second inlet so that light based on the optical statewithin the first reaction vessel is transmittable from the firstreaction vessel to the second photoelectric element, via at least thefirst optical fiber and the second inlet; and a fourth measurementposition, in which the fourth end of the second optical fiber isoptically connected with the second photoelectric element via the secondinlet so that light based on the optical state within the secondreaction vessel is transmittable from the second reaction vessel to thesecond photoelectric element, via at least the second optical fiber andthe second inlet.
 3. The optical measurement device of claim 2, whereinthe first photoelectric element is configured to receive and measurelight of a first specific wavelength or wavelength band; wherein thesecond photoelectric element is configured to receive and measure lightof a second specific wavelength or wavelength band; and wherein thefirst specific wavelength or wavelength band is different from thesecond specific wavelength or wavelength band.
 4. The opticalmeasurement device of claim 2, wherein the second inlet is not driven bythe arranging body Y axis transfer mechanism along the firstpredetermined path.
 5. The optical measurement device of claim 1,wherein the first measurement device further includes a firstirradiation source and a first outlet optically connected with the firstirradiation source; and wherein the optical measurement device furthercomprises: a third optical fiber including opposing fifth and sixthends; and a fourth optical fiber including opposing seventh and eighthends, wherein the fifth end of the third optical fiber and the seventhend of the fourth optical fiber are coupled to the light guide plate,and the light guide stage is driven in the vertical direction relativeto the openings of the first and second reaction vessels by the stage Zaxis transfer mechanism to simultaneously optically connect: the firstend of the first optical fiber and the fifth end of the third opticalfiber with the first interior of the first reaction vessel, and thethird end of the second optical fiber and the seventh end of the fourthoptical fiber with the second interior of the second reaction vessel,wherein the connecting end arranging body supports the sixth end of thethird optical fiber and the eighth end of the fourth optical fiber alonga second predetermined path; and wherein the connecting end arrangingbody is driven by the controller along the second predetermined path bythe arranging body Y axis transfer mechanism between: a first excitationposition, in which the sixth end of the third optical fiber is opticallyconnected with the first irradiation source via the first outlet so thatexcitation light is transmittable from the first irradiation source tothe first reaction vessel, via at least the first outlet and the thirdoptical fiber; and a second excitation position, in which the eighth endof the fourth optical fiber is optically connected with the firstirradiation source via the first outlet so that excitation light istransmittable from the first irradiation source to the second reactionvessel, via at least the first outlet and the fourth optical fiber. 6.The optical measurement device of claim 5, wherein the first measurementposition and the first excitation position coincide with each other sothat, when the second end of the first optical fiber is opticallyconnected with the first photoelectric element via the first inlet, thesixth end of the third optical fiber is optically connected with thefirst irradiation source via the first outlet; and wherein the secondmeasurement position and the second excitation position coincide witheach other so that, when the fourth end of the second optical fiber isoptically connected with the first photoelectric element via the firstinlet, the eighth end of the fourth optical fiber is optically connectedwith the first irradiation source via the first outlet.
 7. A method foroperating an optical measurement device, comprising: providing a firstoptical fiber and a second optical fiber, wherein the first opticalfiber includes opposing first and second ends, and the second opticalfiber includes opposing third and fourth ends; providing a firstreaction vessel and a second reaction vessel, wherein the first reactionvessel comprises an opening to a first interior, and the second reactionvessel comprises an opening to a second interior; providing a firstmeasurement device for receiving emissions from the first and secondreaction vessels, wherein the first measurement device comprises a firstphotoelectric element and a first inlet optically connected with thefirst photoelectric element; driving a light guide stage in a verticaldirection relative to the openings of the first and second interiors,wherein the light guide stage comprises a light guide plate coupled tothe first end of the first optical fiber and the third end of the secondoptical fiber; simultaneously optically connecting the first end of thefirst optical fiber with the first interior of the first reactionvessel, and the third end of the second optical fiber with the secondinterior of the second reaction vessel; driving a connecting endarranging body along a first predetermined path to a first measurementposition, wherein the connecting end arranging body supports the secondend of the first optical fiber and the fourth end of the second opticalfiber along the first predetermined path; optically connecting thesecond end of the first optical fiber with the first photoelectricelement via the first inlet so that light based on an optical statewithin the first reaction vessel is transmitted from the first reactionvessel to the first photoelectric element; driving the connecting endarranging body along the first predetermined path to a secondmeasurement position; and optically connecting the fourth end of thesecond optical fiber with the first photoelectric element so that lightbased on an optical state within the second reaction vessel istransmitted from the second reaction vessel to the first photoelectricelement.
 8. The method of claim 7, further comprising: providing asecond measurement device including a second photoelectric element and asecond inlet optically connected with the second photoelectric element;driving the connecting end arranging body along the first predeterminedpath to a third measurement position; optically connecting the secondend of the first optical fiber with the second photoelectric element viathe second inlet so that light based on the optical state within thefirst reaction vessel is transmitted from the first reaction vessel tothe second photoelectric element; driving the connecting end arrangingbody along the first predetermined path to a fourth measurementposition; and optically connecting the fourth end of the second opticalfiber with the second photoelectric element via the second inlet so thatlight based on the optical state within the second reaction vessel istransmitted from the second reaction vessel to the second photoelectricelement.
 9. The method of claim 8, wherein: the first photoelectricelement is configured to receive and measure light of a first specificwavelength or wavelength band; the second photoelectric element isconfigured to receive and measure light of a second specific wavelengthor wavelength band; and the first specific wavelength or wavelength bandis different from the second specific wavelength or wavelength band. 10.The method of claim 7, further comprising: providing a third opticalfiber and a fourth optical fiber, wherein the third optical fiberincludes opposing fifth and sixth ends, and the fourth optical fiberincludes opposing seventh and eighth ends; wherein the fifth end of thethird optical fiber and the seventh end of the fourth optical fiber arecoupled to the light guide plate; and wherein the connecting endarranging body supports the sixth end of the third optical fiber and theeighth end of the fourth optical fiber along a second predeterminedpath.
 11. The method of claim 10, further comprising: simultaneouslyoptically connecting the fifth end of the third optical fiber with thefirst interior of the first reaction vessel and the seventh end of thefourth optical fiber with the second interior of the second reactionvessel.
 12. The method of claim 11, wherein the first measurement devicefurther comprises a first irradiation source and a first outletoptically connected with the first irradiation source.
 13. The method ofclaim 12, further comprising: driving the connecting end arranging bodyalong the second predetermined path to a first excitation position;optically connecting the sixth end of the third optical fiber with thefirst irradiation source via the first outlet so that excitation lightis transmitted from the first irradiation source to the first reactionvessel; driving the connecting end arranging body along the secondpredetermined path to a second excitation position; and opticallyconnecting the eighth end of the fourth optical fiber with the firstirradiation source via the first outlet so that excitation light istransmitted from the first irradiation source to the second reactionvessel.
 14. The method of claim 13, wherein: the first measurementposition and the first excitation position coincide with each other sothat, when the second end of the first optical fiber is opticallyconnected with the first photoelectric element via the first inlet, thesixth end of the third optical fiber is optically connected with thefirst irradiation source via the first outlet; and the secondmeasurement position and the second excitation position coincide witheach other so that, when the fourth end of the second optical fiber isoptically connected with the first photoelectric element via the firstinlet, the eighth end of the fourth optical fiber is optically connectedwith the first irradiation source via the first outlet.
 15. An opticalmeasurement device, comprising: a plurality of optical fibers, eachoptical fiber having a reaction vessel end and a measurement end; aplurality of reaction vessels, each of the reaction vessels comprisingan opening to an interior; a controller; a light guide stage operativelyconnected to the controller and coupled to the plurality of reactionvessel ends of the optical fibers in a first linear array pattern; aconnecting end arranging body operatively connected to the controllerand coupled to the plurality of measurement ends of the optical fibersin a second linear array pattern; a plurality of measurement devices forreceiving emissions from the plurality of reaction vessels, each of themeasurement devices comprising a photoelectric element and an inletoptically connected with the photoelectric element; wherein thecontroller is configured to: drive the light guide stage in a firstdirection relative to the plurality of openings of the reaction vesselsto simultaneously optically connect the plurality of reaction vesselends of the optical fibers with the plurality of interiors of thereaction vessels; and drive the connecting end arranging body in asecond direction relative to the plurality of inlets of the measurementdevices to sequentially optically connect the plurality of measurementends of the optical fibers with the plurality of photoelectric elementsof the measurement devices such that light based on an optical stateassociated with each of the plurality of reaction vessels istransmittable from the plurality of reaction vessels to the plurality ofphotoelectric elements of the measurement devices.
 16. The opticalmeasurement device of claim 15, wherein the first direction isorthogonal to the second direction.
 17. The optical measurement deviceof claim 15, wherein a pitch of the plurality of reaction vessel ends ofthe optical fibers in the first linear array pattern is greater than apitch of the plurality of measurement ends of the optical fibers in thesecond linear array pattern.
 18. The optical measurement device of claim17, wherein a speed of the connecting end arranging body in the seconddirection relative to the plurality of inlets of the measurement devicesis based on at least one of a stable light receivable time, a lifetimeof fluorescent light with respect to excitation light irradiation, anumber of the plurality of optical fibers, and the pitch of theplurality of measurement ends of the optical fibers in the second lineararray pattern.
 19. The optical measurement device of claim 15, furthercomprising: a nozzle head coupled to and movable with the light guidestage, comprising: a suction-discharge mechanism; and a plurality ofnozzles connected to the suction-discharge mechanism, each of theplurality of nozzles comprising a detachable dispensing tip; whereinsuction and discharge of gases by the suction-discharge mechanism causessuction and discharge of liquids by the plurality of dispensing tips.20. The optical measurement device of claim 19, further comprising: aplurality of transparent sealing lids configured to be coupled to theplurality of nozzles; wherein the controller is further configured tomount each of the plurality of transparent sealing lids on each of theplurality of reaction vessels by detaching the plurality of transparentlids from the plurality of nozzles using a detaching mechanism.